Science.gov

Sample records for agglutinated rabbit erythrocytes

  1. Human rotavirus detection by agglutination of antibody-coated erythrocytes.

    PubMed

    Sanekata, T; Okada, H

    1983-06-01

    We sensitized sheep erythrocytes (SRBC) with antibodies against human rotavirus strain Wa (SRBC-antiWa) and antibodies against calf rotavirus strain NCDV (SRBC-antiNCDV). These were readily agglutinated in the presence of homologous antigens, i.e., human rotavirus and calf rotavirus. By the hemagglutination of SRBC-antiWa and SRBC-antiNCDV (reverse passive hemagglutination [RPHA]), titration of rotavirus in extracts from feces of children suffering from diarrhea (61 specimens) was carried out. We found that the ratio of titers determined with SRBC-antiWa and SRBC-antiNCDV varied remarkably from specimen to specimen. This indicated that the antigenic determinants on human rotavirus in patients feces cross-react with antibodies against NCDV to varying extents. To express the cross-reactivity of human rotavirus with antibodies to NCDV, we propose a Wa/NCDV rotavirus index which can be calculated from the RPHA titer with SRBC-antiWa and SRBC-antiNCDV as follows: Wa/NCDV rotavirus index = (antiWa-RPHA titer of specimen/antiWa-RPHA titer of NCDV)/(antiNCDV-RPHA titer of specimen/antiNCDV-RPHA titer of NCDV).

  2. An inhibitor to erythrocyte agglutination in bovine albumin preparations.

    PubMed

    Gunson, H H; Phillips, P K

    1975-01-01

    Three out of 28 commercial preparations of bovine serum albumin have been encountered which have an inhibitory effect on the assay of anti-Rh-o(D) using the Technicon AutoAnalyser. The inhibitory property, which can also be demonstrated by standard manual serological techniques, appears to be directed towards the second stage of the agglutination reaction. An automated screening procedure for bovine serum albumin preparations and some properties of the inhibitor are described.

  3. Rheologic characterization of vegetal lectins by dissociation of induced erythrocyte agglutinates.

    PubMed

    Rasia, R J; Valverde, J R; Gentils, M; Cauchois, C; Stoltz, J F

    1997-01-01

    Energy evolved from hemagglutination reaction or spent in dissociating erythrocyte agglutinates has been proved to be an excellent parameter for analyzing cell-cell interactions mediated by bridging molecules such as antibodies or lectins. We developed a new rheo-optical method to estimate the energy of dissociation of red blood cell agglutinates. In a Couette shear field agglutinates can be dissociated until a suspension of monodispersed cells is obtained. Intensity of light backscattered by suspended agglutinates increases during their mechanical dissociation. Variation of backscattered light intensity correlates with the energy spent in the process. The adhesive energy of erythrocyte agglutination induced by lectins has been estimated by applying this method. Two specific lectins (Dolichus Biflorus agglutinin and Ulex Europaeus agglutinin) and a new lectin obtained from Amarantus Cruentus seeds which specificity is unknown were studied. Results obtained in this work for Dolichus Biflorus lectin are comparable with values published by other authors. An asymptotic decrease of adhesive energy was observed when the mechanical dissociation was applied several times on the same sample. Our results suggest that the cell detachment is accompanied by the extraction of membrane receptors. This finding is consistent with results obtained by other authors.

  4. 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. PMID:2066386

  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. 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

  9. 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.

  10. Comparative evaluation of gel column agglutination and erythrocyte magnetized technology for red blood cell alloantibody titration.

    PubMed

    Dubey, Anju; Sonker, Atul; Chaudhary, Rajendra K

    2015-01-01

    Antibody titration is traditionally performed using a conventional test tube (CTT) method, which is subjected to interlaboratory variations because of a lack of standardization and reproducibility. The aim of this study is to compare newer methods such as get column technology (GCT) and erythrocyte magnetized technology (EMT) for antibody titration in terms of accuracy and precision. Patient serum samples that contained immunoglobin G (IgG) red blood cell (RBC) alloantibodies of a single specificity for Rh or K anitgens were identified during routine transfusion service testing and stored. Titration and scoring were performed separately by and stored. Titration and scoring were performed separately by different laboratory personnel on CTT, GCT, and EMT. Testing was performed a total of three times on each sample. Results were analyzed for accuracy and precision. A total of 50 samples were tested. Only 20 percent of samples tested with GCT shoed titers identical to CTT, whereas 48 percent of samples tested with EMT showed titers identical to CTT. Overall, the mean of th titer difference from CTT was higher using GCT (+0.31) compared with that using EMT (+0.13). Precision shown by CTT was 30 percent, EMT was 76 percent, and GCT was 92 percent on repeat testing. GCT showed higher titer values in comparison with CTT but was found to be the most precise. EMT titers were comparable to CTT, and its precision was intermediate. Further studies to validate this method are required.

  11. Evidence for sialylated type 1 blood group chains on human erythrocyte membranes revealed by agglutination of neuraminidase-treated erythrocytes with Waldenström's macroglobulin IgMWOO and hybridoma antibody FC 10.2.

    PubMed

    Picard, J K; Loveday, D; Feizi, T

    1985-01-01

    Haemagglutination studies have been performed with untreated and neuraminidase-treated human erythrocytes of the three Lewis antigen types Le(a-b-), Le(a+b-) and Le(a-b+) using two monoclonal antibodies, IgMWOO and FC 10.2, which were previously shown to recognize the type 1 based blood group chains: Gal beta 1----3GlcNAc beta 1----3Gal beta 1----4Glc/GlcNAc (for explanation of abbreviations see table IV legend). Both antibodies behaved as cold agglutinins with neuraminidase-treated but not with untreated erythrocytes of the three Lewis antigen types. Neuraminidase-treated erythrocytes of i antigen type were similarly agglutinated. This haemagglutination was specifically inhibited by the type 1 based milk oligosaccharide lacto-N-tetraose. Thus, there is strong evidence for the occurrence of sialylated type 1 chains on human erythrocyte membranes of I and i antigen types. In addition, evidence for the presence of type 1 chains which are both sialylated and fucosylated was obtained by (1) haemagglutination of Le(a+b-) erythrocytes with the monoclonal antibody 19.9; (2) increased haemagglutination of neuraminidase-treated erythrocytes with anti-H antibodies of Bombay serum; (3) increased haemagglutination of neuraminidase-treated Le(a+b-) cells with anti-Lea antibodies, and (4) the appearance of Lea antigen activity on neuraminidase-treated erythrocytes of Le(a-b+) type.

  12. 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.

  13. Toxin a from Clostridium difficile binds to rabbit erythrocyte glycolipids with therminal Gal. cap alpha. 1-3Gal. beta. 1-4GlcNaC sequences

    SciTech Connect

    Clark, G.F.; Krivan, H.; Wilkins, T.; Smith, D.F.

    1987-05-01

    Toxin A is one of two clostridial toxins implicated as the causative agent of pseudomembranous colitis in patients undergoing postoperative antibiotic therapy. Evidence that the carbohydrate binding determinant for this toxin is a glycoconjugate(s) with non-reducing Gal..cap alpha..1-3Gal..beta..1-4GlcNAc has recently been reported. Specific agglutination of rabbit erythrocytes by Toxin A is inhibited by bovine thyroglobulin and prevented by pretreatment of cells with ..cap alpha..-galactosidase. Total lipid extracts from rabbit erythrocytes were subjected to thin layer chromatography and the chromatogram overlaid with purified /sup 125/I-labeled Toxin A. Two major and several minor toxin-binding glycolipids were detected following autoradiography. The major toxin-binding glycolipids were identified as pentasaccharide- and decasaccharide-ceramides expressing terminal Gal..cap alpha..1-3Gal..beta..1-4GlcNAc sequences. Treatment of the toxin-binding glycolipids with ..cap alpha..-galactosidase abolished binding. Forsmann glycolipid, globoside, Gal..cap alpha..1-4 Gal..beta..1-4Glc-cer, and Gal..cap alpha..1-3Gal..beta..1-4Glc-cer did not bind the toxin. These observations are consistent with the proposed carbohydrate specificity of the toxin for the non-reducing terminal sequence, Gal..cap alpha..1-3Gal..beta..1-4GlcNAc.

  14. Free radicals promote in vitro intracellular decay of rabbit reticulocyte and erythrocyte hexokinase

    SciTech Connect

    Stocchi, V.; Biagiarelli, B.; Masat, L.; Palma, F.; Piccoli, G.; Cucchiarini, L. )

    1991-03-15

    The authors studied the behavior of enzymes involved in the glycolytic pathway incubating intact reticulocyte and erythrocyte at 37C in the presence of ascorbic acid and Fe{sup 2+}. The results obtained have shown evidence that among the glycolytic enzymes the hexokinase activity shows a pronounced decay. For this reason the authors have investigated how the chromatographic profile of hexokinase changes after exposure of reticulocytes and erythrocytes to the oxygen-radical generating system in trying to understand the molecular basis of this inactivation. The results obtained have shown a different effect of free radicals on the reticulocyte and erythrocyte hexokinase molecular forms. The analysis of the chromatographic profile was performed using a TSK Gel Toyopearl DEAE 650 S column which allows a complete resolution of the distinct forms of hexokinase together with a complete recovery of the enzyme activities. Concomitantly to the hexokinase decay, there is a fall in the GSH level when intact rabbit erythrocytes and reticulocytes are incubated in presence of iron and ascorbic acid. However, the fall of GSH is significantly higher in the erythrocytes where, after incubation of one hour, it reaches a mean value of 0.3 {mu}moles per ml of cells representing about 10% of the initial value. In the reticulocytes the GSH value, after the same treatment, remain as high as 1.6 {mu}moles per ml of cells. If we consider that the initial value of GSH is almost the same in the erythrocytes are reticulocytes the highest decay rate of hexokinase observed in these latter cells, cannot be related to the fall of the GSH level, but to a possible direct effect of free-radicals on the enzyme.

  15. HEMOLYSIS OF RABBIT ERYTHROCYTES BY PURIFIED STAPHYLOCOCCAL ALPHA-TOXIN. I. KINETICS OF THE LYTIC REACTION.

    PubMed

    COOPER, L Z; MADOFF, M A; WEINSTEIN, L

    1964-01-01

    Cooper, Louis Z. (New England Center Hospital, Boston, Mass.), Morton A. Madoff, and Louis Weinstein. Hemolysis of rabbit erythrocytes by purified staphylococcal alpha-toxin: I. Kinetics of the lytic reaction. J. Bacteriol. 87:127-135. 1964.-The hemolytic activity of purified staphylococcal alpha-lysin was found to be directly proportional to toxin concentration and inversely related to the log concentration of rabbit erythrocytes. Activity was directly proportional to the duration of lysin-red cell incubation until inactivating effects of heat and dilution became significant; this linear relationship was prolonged by incubation at a lower temperature and addition of bovine serum albumin. Study of the time course of hemolysis at different alpha-lysin concentrations revealed a family of sigmoid curves characterized by a prelytic lag phase and a period of rapid linear release of hemoglobin. The duration of prelytic lag varied inversely with the quantity of toxin, but the rate of hemolysis was directly proportional to toxin and red-cell concentrations. The presence of bovine serum albumin decreased the prelytic lag, prolonged the linear phase of the reaction, and increased total hemolysis. In the range of 25 to 46 C, the prelytic lag period became shorter with increase in temperature; at 48 to 52 C, it was markedly prolonged and hemolysis was strikingly diminished. As the incubation temperature was increased from 25 to 52 C, there was a decrease in the degree of maximal hemolysis, presumably due to thermal inactivation of alphalysin. The rate of hemolysis, when measured to 50% hemolysis, was optimal between 34 and 42 C but, when determined to the 10% level, was greatest between 40 and 46 C. The features of the hemolytic reaction suggest that staphylococcal alpha-toxin has the characteristics of an enzyme.

  16. Iloprost- and isoproterenol-induced increases in cAMP are regulated by different phosphodiesterases in erythrocytes of both rabbits and humans.

    PubMed

    Adderley, Shaquria P; Dufaux, Eileen A; Sridharan, Meera; Bowles, Elizabeth A; Hanson, Madelyn S; Stephenson, Alan H; Ellsworth, Mary L; Sprague, Randy S

    2009-05-01

    Activation of the G protein G(s) results in increases in cAMP, a necessary step in the pathway for ATP release from rabbit and human erythrocytes. In all cells, the level of cAMP is the product of its synthesis by adenylyl cyclase and its hydrolysis by phosphodiesterases (PDEs). Both iloprost (Ilo), a PGI(2) analog, and isoproterenol (Iso), a beta-agonist, stimulate receptor-mediated increases in cAMP in rabbit and human erythrocytes. However, the specific PDEs associated with each of these signaling pathways in the erythrocyte have not been fully characterized. Previously, we reported that PDE3B is present in rabbit and human erythrocyte membranes and that PDE3 inhibitors potentiate Ilo-induced increases in cAMP. Here we report that inhibitors of either PDE2 or PDE4, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and rolipram, respectively, potentiate Iso-induced increases in cAMP in rabbit and human erythrocytes. Importantly, these inhibitors had no effect on cAMP increases associated with the incubation of erythrocytes with Ilo. In addition, we establish, for the first time, the presence of PDE2A protein in rabbit and human erythrocyte membranes. Finally, we determined that preincubation of human erythrocytes with EHNA and rolipram together potentiate Iso-induced ATP release, whereas preincubation with cilostazol enhances Ilo-induced release of ATP. These results are consistent with the hypothesis that, in rabbit and human erythrocytes, Ilo-induced increases in cAMP and ATP release are regulated by PDE3, whereas those associated with Iso are regulated by the activities of both PDE2 and PDE4. These studies demonstrate that PDE activity in these cells is localized to specific signaling pathways. PMID:19252089

  17. Studies on the role of goat heart galectin-1 as an erythrocyte membrane perturbing agent.

    PubMed

    Ashraf, Ghulam Md; Perveen, Asma; Zaidi, Syed Kashif; Tabrez, Shams; Kamal, Mohammad A; Banu, Naheed

    2015-01-01

    Galectins are β-galactoside binding lectins with a potential hemolytic role on erythrocyte membrane integrity and permeability. In the present study, goat heart galectin-1 (GHG-1) was purified and investigated for its hemolytic actions on erythrocyte membrane. When exposed to various saccharides, lactose and sucrose provided maximum protection against hemolysis, while glucose and galactose provided lesser protection against hemolysis. GHG-1 agglutinated erythrocytes were found to be significantly hemolyzed in comparison with unagglutinated erythrocytes. A concentration dependent rise in the hemolysis of trypsinized rabbit erythrocytes was observed in the presence of GHG-1. Similarly, a temperature dependent gradual increase in percent hemolysis was observed in GHG-1 agglutinated erythrocytes as compared to negligible hemolysis in unagglutinated cells. The hemolysis of GHG-1 treated erythrocytes showed a sharp rise with the increasing pH up to 7.5 which became constant till pH 9.5. The extent of erythrocyte hemolysis increased with the increase in the incubation period, with maximum hemolysis after 5 h of incubation. The results of this study establish the ability of galectins as a potential hemolytic agent of erythrocyte membrane, which in turn opens an interesting avenue in the field of proteomics and glycobiology.

  18. Toxin A from Clostridium difficile binds to rabbit erythrocyte glycolipids with terminal Gal alpha 1-3Gal beta 1-4GlcNAc sequences

    SciTech Connect

    Clark, G.F.; Krivan, H.C.; Wilkins, T.D.; Smith, D.F.

    1987-08-15

    The binding of Toxin A isolated from Clostridium difficile to rabbit erythrocyte glycolipids has been studied. Total lipid extracts from rabbit erythrocytes were subjected to thin-layer chromatography and toxin-binding glycolipids detected by using /sup 125/I-labeled Toxin A in a direct binding overlay technique. Two major and several minor toxin-binding glycolipids were detected in rabbit erythrocytes by this method. The results of structural analyses of the major toxin-binding glycolipids were consistent with a pentasaccharide-ceramide (Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc-Cer) and a branched decasaccharide-ceramide (Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-3(Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-6)Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc-Cer) previously identified as the two most abundant glycolipids in rabbit erythrocytes. /sup 125/I-Toxin A binding to these glycolipids could be inhibited by bovine thyroglobulin, monospecific antiserum to the toxin, or by treatment of the glycolipids with alpha-galactosidase. The absence of toxin interaction with isoglobotriaosylceramide (Gal alpha 1-3Gal beta 1-4Glc-Cer) isolated from canine intestine suggested that the GlcNAc residue present in the terminal Gal alpha 1-3Gal beta 1-4GLcNAc sequence common to all known toxin binding glycoconjugates is required for carbohydrate-specific recognition by Toxin A. These observations are consistent with the proposed carbohydrate binding specificity of Toxin A for the nonreducing terminal sequence, Gal alpha 1-3Gal beta 1-4GlcNAc.

  19. Binding of Rabbit Hemorrhagic Disease Virus to Antigens of the ABH Histo-Blood Group Family

    PubMed Central

    Ruvoën-Clouet, Nathalie; Ganière, Jean Pierre; André-Fontaine, Geneviève; Blanchard, Dominique; Le Pendu, Jacques

    2000-01-01

    The ability of rabbit hemorrhagic disease virus to agglutinate human erythrocytes and to attach to rabbit epithelial cells of the upper respiratory and digestive tracts was shown to depend on the presence of ABH blood group antigens. Indeed, agglutination was inhibited by saliva from secretor individuals but not from nonsecretors, the latter being devoid of H antigen. In addition, erythrocytes of the rare Bombay phenotype, which completely lack ABH antigens, were not agglutinated. Native viral particles from extracts of infected rabbit liver as well as virus-like particles from the recombinant virus capsid protein specifically bound to synthetic A and H type 2 blood group oligosaccharides. Both types of particles could attach to adult rabbit epithelial cells of the upper respiratory and digestive tracts. This binding paralleled that of anti-H type 2 blood group reagents and was inhibited by the H type 2-specific lectin UEA-I and polyacrylamide-conjugated H type 2 trisaccharide. Young rabbit tissues were almost devoid of A and H type 2 antigens, and only very weak binding of virus particles could be obtained on these tissues. PMID:11090195

  20. 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.

  1. Sm10.3, a Member of the Micro-Exon Gene 4 (MEG-4) Family, Induces Erythrocyte Agglutination In Vitro and Partially Protects Vaccinated Mice against Schistosoma mansoni Infection

    PubMed Central

    Martins, Vicente P.; Morais, Suellen B.; Pinheiro, Carina S.; Assis, Natan R. G.; Figueiredo, Barbara C. P.; Ricci, Natasha D.; Alves-Silva, Juliana; Caliari, Marcelo V.; Oliveira, Sergio C.

    2014-01-01

    Background The parasitic flatworm Schistosoma mansoni is a blood fluke that causes schistosomiasis. Current schistosomiasis control strategies are mainly based on chemotherapy, but many researchers believe that the best long-term strategy to control disease is a combination of drug treatment and immunization with an anti-schistosome vaccine. Numerous antigens that are expressed at the interface between the parasite and the mammalian host have been assessed. Among the most promising molecules are the proteins present in the tegument and digestive tract of the parasite. Methodology/Principal Findings In this study, we evaluated the potential of Sm10.3, a member of the micro-exon gene 4 (MEG-4) family, for use as part of a recombinant vaccine. We confirmed by real-time PCR that Sm10.3 was expressed at all stages of the parasite life cycle. The localization of Sm10.3 on the surface and lumen of the esophageal and intestinal tract in adult worms and lung-stage schistosomula was confirmed by confocal microscopy. We also show preliminary evidence that rSm10.3 induces erythrocyte agglutination in vitro. Immunization of mice with rSm10.3 induced a mixed Th1/Th2-type response, as IFN-γ, TNF-α, and low levels of IL-5 were detected in the supernatant of cultured splenocytes. The protective effect conferred by vaccination with rSm10.3 was demonstrated by 25.5–32% reduction in the worm burden, 32.9–43.6% reduction in the number of eggs per gram of hepatic tissue, a 23.8% reduction in the number of granulomas, an 11.8% reduction in the area of the granulomas and a 39.8% reduction in granuloma fibrosis. Conclusions/Significance Our data suggest that Sm10.3 is a potential candidate for use in developing a multi-antigen vaccine to control schistosomiasis and provide the first evidence for a possible role for Sm10.3 in the blood feeding process. PMID:24651069

  2. Effect of garlic supplementation on erythrocytes antioxidant parameters, lipid peroxidation, and atherosclerotic plaque formation process in oxidized oil-fed rabbits.

    PubMed

    Zalejska-Fiolka, Jolanta; Kasperczyk, Aleksandra; Kasperczyk, Sławomir; Błaszczyk, Urszula; Birkner, Ewa

    2007-01-01

    Effect of garlic supplementation on blood antioxidant status, lipid peroxidation, and coronary plaque formation process was investigated in oxidized oil-fed rabbits. Eighteen adult male mixed European rabbits were given a balanced diet (21 g% protein, 34 g% fat, 45 g% carbohydrate), which contained isocaloristic addition of nonoxidized or oxidized rapeseed oil in the presence and absence of garlic. The experiment lasted 24 weeks. At the beginning and every 6 weeks, rabbits were weighed, and blood was taken. To evaluate the antioxidant status of the rabbits, erythrocytes malondialdehyde (MDA) concentration, total superoxide dismutase (t-SOD), and glutathione peroxidase (GPX) activations were determined. After the experiment was completed, aortas were dissected for histological examinations. Changes in the contents of the above parameters and histological examinations showed that oxidized rapeseed, oil administered to rabbits, caused the development of atherosclerotic changes and disturbed antioxidant status. The addition of garlic in such diets inhibited atherosclerotic changes in the aorta wall, and it is related to the homeostatic activity of antioxidative enzymes and lipid peroxidation.

  3. [Na+/H+- and Na+/Na+-countertransport in human, rabbit, and rat erythrocytes: evidence for the existence of two independent ion-transporting systems].

    PubMed

    Orlov, S N; Kuznetsov, S R; Kolosova, I A; Makarov, V L

    1994-05-01

    The activity and regulatory features of the Na+/H(+)- and Na+/Na(+)-exchange were studied in human, rabbit and rat red blood cells. No basal activity of the Na+/H(+)-exchange (the amyloride-inhibited component of the 22Na+ influx) in erythrocytes of these species was observed. The rate of 22Na+ influx increased rapidly when the experiments were carried out on acid-loaded cells in an alkaline (pH0 = 8.0) incubation medium (delta mu H(+)-induced Na+/H(+)-exchange). The ratio of delta mu H(+)-induced Na+/H(+)-exchange activities in human, rabbit and rat red blood cells was 1.0 : 1.1 : 2.3, respectively, whereas that of the Na+/Na(+)-exchange activities (the phloretin-inhibited component of the 22Na+ influx) in erythrocytes of these species was 1.0 : 4.6 : 0.2. The osmotic shrinkage of rat and rabbit erythrocytes led to the stimulation of the Na+/H(+)- (but not Na+/Na+) exchange. Amyloride (1 mM) inhibited the shrinkage-induced 22Na+ entry as well as the delta mu H(+)-induced 22Na+ entry--by 95 and 10-20%, respectively. Heat treatment (10 min, 49-51 degrees C), disturbing the membrane cytoskeleton suppressed both the shrinkage-induced activation and the delta mu H(+)-induced activation of the Na+/H(+)-exchange. The data obtained indicate that the both transport systems are mediated by two distinct transport carriers. It may be suggested that the delta mu H(+)-induced Na+/H(+)-exchange, on the one hand, and the shrinkage-induced Na+/H(+)-exchange, on the other, are mediated by two different Na+/H(+)-exchanger subtypes. PMID:8043690

  4. 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

  5. Erythrocyte 2,3-diphosphoglycerate and erythropoietic activity in rabbits with severe bleeding anaemia superimposed in the early post-natal fall in haemoglobin.

    PubMed

    Holter, P H; Refsum, H E

    1985-08-01

    Erythrocyte 2,3-diphosphoglycerate (2,3-DPG), whole blood haemoglobin (Hb), haematocrit (Hct), mean corpuscular haemoglobin concentration (MCHC) and reticulocyte percentage (Rt) were determined before and after bleeding in two groups of suckling chinchilla rabbits. One group was subjected to severe bleeding on the 12th, 15th, 18th and 21st day after birth and studied for 3 weeks, while the other was studied during the first 24 h after one severe bleeding on the 18th day. In the first group Hb and Hct fell to 2.7 g . dl-1 and 11.5%, respectively, on the 25th day. The fall was accompanied by a marked rise in Rt and decline in MCHC, reaching maximum and minimum on the 29th day, and a simultaneous, temporary decline in 2,3-DPG. In the other group the acute bleeding was accompanied by a marked fall in Hb and Hct, but no change in Rt and MCHC. 2,3-DPG was unchanged 8 h after the bleeding, but showed a rise during the following 16 h, definitely beyond the normal rise at this age. It is concluded that severe bleeding anaemia induces a rise in erythrocyte 2,3-DPG synthesis. However, the effect of the acute rise in 2,3-DPG is far from sufficient to maintain the O2 delivery capacity of the blood, and is completely offset by the influence of a subsequent change in the erythrocyte population towards younger cells with low 2,3-DPG. PMID:4050480

  6. Removal of leucocytes from whole blood and erythrocyte suspensions by filtration through cotton wool. IV. Immunization studies in rabbits.

    PubMed

    Engelfriet, C P; Diepenhorst, P; Van den Giessen, M; von Riesz, E

    1975-01-01

    Hetero-immunization experiments in rabbits were performed to evaluate the quantity of leucocyte antigens present in red cell suspensions prepared by the filtration method of DIEPENHORST et al., in which no intact leucocytes could be detected. It was found that the quantity of leucocyte antigens is smallest in blood filtered immediately after taking. More leucocyte-antigenic material seems to be present in blood that is filtered after storage and in frozen-thawed red cell suspensions. Evidence was obtained that granulocyte-specific antigens are more strongly immunogenic in the rabbit than lymphocyte-specific antigens or antigens common to both cells.

  7. 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

  8. ERYTHROCYTE SENSITIZATION BY BLOOD GROUP-SPECIFIC BACTERIAL ANTIGENS.

    PubMed

    SPRINGER, G F; HORTON, R E

    1964-07-01

    Human and chicken erythrocytes are readily coated in vitro by blood group active protein-lipopolysaccharides and lipopolysaccharides from E. coli O(86) and E. coli O(128). Serum albumin, alpha(2)- and beta-lipoproteins inhibit this sensitization. Blood group B specific agglutination of erythrocytes with B or B-like antigens was obtained with antibodies purified by adsorption on and elution from B erythrocytes. Anti-blood group B and E. coli O(86)-specific antibodies could be eluted from E. coli O(86)-coated O erythrocytes. Eel anti-H(O) serum agglutinated O erythrocytes and only those A(1)B red cells which were coated with blood group H(O) active E. coli products. Blood group active substances specifically inhibited agglutination of lipopolysaccharide-coated erythrocytes by anti-B and anti-H(O) agglutinins. Demonstrable amounts of lipopolysaccharide could only be removed from coated erythrocytes by washing them at elevated temperatures (58 degrees C) in physiological solutions. Red cell sensitization with B active E. coli O(86) substances was achieved in vivo in a minority of severely diseased infants and in germ-free and ordinary chicks which were in tourniquet shock after treatment with cathartics. Therefore, a possible mode by which erythrocytes of patients with severe intestinal disorders acquire antigens is the fixation of bacterial substances to their surfaces, if there are not enough of the normally interfering plasma factors present.

  9. Membrane proteins in senescent erythrocytes.

    PubMed Central

    Suzuki, T; Dale, G L

    1989-01-01

    The examination of erythrocyte senescence has been facilitated by recent advances in techniques for the isolation of aged red cells. One of these methods, which uses biotinylated rabbit erythrocytes, has been used to examine the state of membrane proteins in effete cells. These aged red cells were found to have normal ratios of alpha-spectrin and beta-spectrin as well as normal levels of ankyrin. The observation concerning ankyrin is particularly important due to the sensitivity of this protein to proteolysis and the postulated action of proteinases in the aging process. The senescent erythrocytes were also found to have an altered ratio of bands 4.1a and 4.1b without any apparent change in the total level of 4.1. In addition, the analysis of the aged cell membranes did not show any large-molecular-mass aggregated protein at the origin of the SDS/polyacrylamide gels, indicating a lack of transglutaminase activity in the senescence process for rabbit erythrocytes. These results indicate that aging of the rabbit erythrocyte is not accompanied by gross proteolytic degradation or transglutaminase-catalysed cross-linking of membrane components. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. PMID:2522000

  10. 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

  11. Manufacturing High-Fidelity Lunar Agglutinate Simulants

    NASA Technical Reports Server (NTRS)

    Gutafson, R. J.; Edmunson, J. E.; Rickman, D. L.

    2010-01-01

    The lunar regolith is very different from many naturally occurring material on Earth because it forms in the unique, impact-dominated environment of the lunar surface. Lunar regolith is composed of five basic particle types: mineral fragments, pristine crystalline rock fragments, breccia fragments, glasses of various kinds, and agglutinates (glass-bonded aggregates). Agglutinates are abundant in the lunar regolith, especially in mature regoliths where they can be the dominant component.This presentation will discuss the technical feasibility of manufacturing-simulated agglutinate particles that match many of the unique properties of lunar agglutinates.

  12. Protein-sugar interactions: preparation, purification, and properties of rabbit antibodies against di-N-acetylchitobiose.

    PubMed Central

    Kieda, C M; Delmotte, F M; Monsigny, M L

    1977-01-01

    Antibodies against di-N-acetylchitobiose (CB) were raised in rabbits after injection of CB-bovine serum albumin conjugates and were fractionated by columns into two classes: the first bound to a column of Sepharose covalently coupled with N-acetylglucosamine (GlcNAc); the second bound to a column coupled with CB. Active antibodies were eluted by a moderate concentration of a chaotropic agent, but not by high ionic strength buffers or acidic buffers. The active fractions were identified as IgG by ultracentrifugation and immuno-electrophoresis. These antibodies gave precipitation bands with CB-protein conjugates and this reaction could be reversed in the presence of free CB. The GlcNAc-bound fraction quantitatively quenched the fluorescence of O-(4-methylumbelliferyl)-glucosides of GlcNAc and CB, while the CB-bound fraction quenched only the glycoside of CB; other O-(4-methylumbelliferyl)-glucosides were not quenched. Among eleven monosaccharides and oligosaccharides, only GlcNAc, CB, and tri-N-acetylchitotriose were able to inhibit the precipitation of antibodies against CB with CB-protein conjugate. These antibodies failed to agglutinate erythrocytes from various species but did agglutinate transformed cells and mouse lymphocytes. The binding of these antibodies on cell lembranes was reversed by free CB and by CB-protein conjugates. The properties of these antibodies are related to those of lectins with similar specificities and to the structure of glycoconjugates. Images PMID:264671

  13. Rapid method for detection of Coxiella burnetii antibodies using high-density particle agglutination.

    PubMed Central

    Nguyen, S V; Otsuka, H; Zhang, G Q; To, H; Yamaguchi, T; Fukushi, H; Noma, A; Hirai, K

    1996-01-01

    A high-density particle agglutination test, using erythrocyte-sensitizing substance from phase II Coxiella burnetii adsorbed to high-density composite particles, was developed for rapid serodiagnosis of Q fever. The test was compared with the microimmunofluorescence test for sensitivity and specificity by using 3,036 human serum samples collected in Gifu Prefecture, Japan. An excellent agreement was found between the two tests for the acute-phase group and paired serum samples, but some discordant results were observed in the single-sample group. The sensitivity and specificity of the high-density particle agglutination test were both 100% in the former group and 81.6 and 99.9%, respectively, in the latter group. The test is a very promising tool for routine serodiagnosis of Q fever because of its simplicity, sensitivity, and specificity. PMID:8940428

  14. 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.

  15. Column agglutination technology: the antiglobulin test.

    PubMed

    Reis, K J; Chachowski, R; Cupido, A; Davies, D; Jakway, J; Setcavage, T M

    1993-08-01

    A new system for typing and screening blood, based on the sieving effect of glass bead microparticles, has been developed. The test is performed in a microcolumn in which the red cell agglutinates are trapped in the glass bead matrix during centrifugation, and unagglutinated cells form a pellet at the bottom of the column. Anti-human globulin reagents were incorporated in the diluent and the new test system, column agglutination technology, was compared to conventional tube tests and low-ionic-strength method. Sera and plasmas (228 samples) were screened for red cell antibodies with two anti-human globulin reagents: one containing only anti-IgG and the other containing both anti-IgG and anti-C3b, -C3d. After initial testing, there was 94-percent agreement between column agglutination technology and tube tests, and after repeat testing, there was 97-percent agreement. The column agglutination technology anti-human globulin test eliminates the need to wash red cells, which decreases the overall test time. The test is easy to perform, and the results are more objective than those with tube and microplate methods.

  16. The production curve for agglutinates in planetary regoliths

    NASA Astrophysics Data System (ADS)

    McKay, D. S.; Basu, A.

    1983-11-01

    Models for the production of agglutinates are developed that can be applied to the lunar surface or to any planetary or asteroidal body lacking an atmosphere. Models are developed using rate equations for progressively more complex situations and range from Model 1, which is a simple linear increase of agglutinate content with time, to Model 4, which includes provision for recycling of existing agglutinates and replenishment and burial of exposed soil. Model 4 has some aspects of a steady state because, depending on the rate constants, agglutinate content may be limited to an intermediate value, even for long exposure times. In an extreme case, agglutinate content may be limited to a value near zero. These models predict that agglutinates should be low in abundance in areas of thin regolith, such as the Lunokhod-2 site on the moon, and on asteroids. The models may also help explain the apparent low agglutinate abundances of lunar regolith breccias and meteorite regolith breccias.

  17. The significance of erythrocyte antigen site density

    PubMed Central

    Hoyer, Leon W.; Trabold, Norma C.

    1970-01-01

    The importance of antigen site density has been studied by means of a model passive hemagglutination system using human red cells coupled with sulfanilic acid groups. Relative site numbers were estimated from the covalent linkage of sulfanilic acid-35S to red cell membrane protein and the effective antigen site number was determined with 125I-labeled rabbit IgG antisulfanilic acid. Cells which had fewer than 20,000 antigen sites per cell were not agglutinated. As greater numbers of sulfanilic groups were coupled to the red cells, the agglutination titers increased to maximum values with red fanilic groups were coupled to the red cells, the agglutination titers of purified IgM antibody were 10-20 times greater than IgG antibody when preparations with the same protein concentration were compared, but this difference was not noted when IgG antibody was measured by antiglobulin reactions. These findings emphasize the need to consider differences in antigen site density when comparing blood group systems. They are consistent with the hypothesis that those blood group antigens which have a very low site number will not be detected by IgG antibodies in saline hemagglutination determinations. PMID:5409811

  18. Functional and structural changes in human erythrocyte surface after irradiation by uv waves of various wavelengths. Report 1: expression of ABO and Rhesus system antigen

    SciTech Connect

    Samoylova, K.A.; Klimova, K.N.; Priyezzheva, L.S.; Artsishevskaya, R.A.

    1985-01-01

    The effect of shortwave ultraviolet (SUV) radiation ad causes change in the external surface of human erythrocytes, modifying the expression of the ABO and Rh system antigens which are related to the surface of the cells was investigated. Erythrocytes in a structurally prepared erythrocyte mass from 23 donors stabilized by glugicir or heparin were examined. Three series of experiments were performed: (1) isolated erythrocytes, before irradiation thrice washed to remove plasma with isotonic NaC1 0.9%, erythrocytes diluted to 5 x s10 to the 7th power cells per milliliter and erythrocytes on the undiluted erythrocyte mass about 7 x 5 x 109 to the 9th power cells power milliliter. The agglutinating activity of the ABO and Rh antigens was studied. Two to three hours after exposure to 248, 620, 1240 and 2480 J/m2, the degree of hemolysis of isolated erythrocytes increased by 5,10,18 and 28%. Changes were also observed in agglutinating activity of ABO antigens. The agglutinating activity of A and B antigens increased by an average factor of 2 minus H antigens by a factor of 4. The SUV radiation did not cause any activation of the Rh antigen.

  19. Agglutinates and carbon accumulation in Apollo 17 lunar soils

    NASA Technical Reports Server (NTRS)

    Basu, A.; Meinschein, W. G.

    1976-01-01

    A critical review of maturity with respect to the abundance of implanted solar wind elements (SWE) in lunar soils indicates: (1) that the Rosiwal Principle has limited applicability in determining implantation of SWE in lunar soils, and (2) that despite a depletion of SWE in agglutinitic glass, agglutinates are enriched in SWE due to the presence of buried surfaces of numerous clasts within agglutinates. A statistical analysis of published data of several Apollo 17 soils indicates that the abundance of carbon and, by analogy, the abundance of other SWE are correlatable with the agglutinate content and the mean grain size of lunar soils. Microscopic examination of more than 5000 grains of agglutinates in polished thin sections reveals a wide range of variability in the mineralogy, grain size distribution, degree of recycling, etc., of the clast population in agglutinates. This indicates that the volume-correlated SWE content of agglutinates may vary and need not be constant.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Is nitrotyrosine generated in human erythrocytes in circulation?

    PubMed

    Kikugawa, K; Nakauchi, K; Beppu, M; Hiramoto, K; Ando, K; Hayakawa, M

    2000-04-01

    Nitrotyrosine is considered a stable biomarker of reactive nitrogen species, including nitrogen dioxide (NO2) and peroxynitrous acid (ONOOH) in biomaterials. There are inconsistent observations on the detection of free and protein-associated nitrotyrosine in normal human plasma. Human erythrocytes, differentiated from erythrocyte precursor cells in the bone marrow, circulating in the body for an average of 120 d, and finally removed by spleen macrophages, may be exposed to reactive nitrogen species. In the present study, membrane proteins and hemoglobin from the senescent erythrocyte population were compared with those from young erythrocytes separated from the same individuals in their nitrotyrosine presence using newly prepared rabbit polyclonal anti-nitrotyrosine-ribonuclease A and anti-nitro(N-butoxycarbonyl)tyrosine-bovine serum albumin antibodies. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the membranes and hemoglobin, and subsequent Western blot analysis, showed that these antibodies only slightly bind to the bands of the proteins from both young and senescent erythrocytes, whereas these antibodies definitely bind to the protein bands of membranes and hemoglobin nitrated by NO2 or ONOOH in vitro. This result indicates that nitrotyrosine is not detected in the membrane proteins and hemoglobin in human normal erythrocytes in circulation. However, this does not conclude that erythrocytes are not exposed to reactive nitrogen species in the circulation.

  6. Diet-induced hypercholesterolaemia in the rabbit.

    PubMed

    Ivanov, A S; Torkhovskaya, T I; Khalilov, E M; Archakov, A I

    1991-01-01

    Intraspecies variation in diet-induced hypercholesterolaemia in rabbits, simulating atherosclerosis, was studied. The chinchilla rabbit population examined contained several subpopulations, as indicated by polymodal forms of histograms of plasma cholesterol levels. This finding indicates that the inclusion of subpopulations in an investigation can lead to erroneous conclusions, and that subpopulations should be identified before such work is undertaken. The relationship between the molar cholesterol/phospholipid ratio in rabbit erythrocyte membranes and the plasma cholesterol level in experimental atherosclerosis was also studied. A correlation was evident only over the range of 1-5 g cholesterol per litre of plasma. PMID:1751762

  7. 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.

  8. 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.

  9. Measuring glass abundances in lunar agglutinates. [Abstract only

    NASA Technical Reports Server (NTRS)

    Strait, M. M.; Basu, A.; Mckay, D. S.; Robinson, R.

    1994-01-01

    The purpose of this study is to develop a standard method for measuring modal abundances of glass in single agglutinates in the lunar regolith. Not only does agglutinitic glass increase in single agglutinates as clasts of older agglutinates get incorporated into newer agglutinates with increasing maturity, but it is in this glassy phase that nanophase superparamagnetic Fe metal originates as a result of reduction reactions during the agglutination process. We report the results of two sets of independent measurements using two different methods to determine the proportion of glass in single agglutinates. We have used polished grain mounts (PGM) of five hand-picked single agglutinates from Apollo 16 soil 61181. The ISI Scanning Electron Microscope (SEM) fitted with a high-resolution Backscattered Electron (BSE) detector was used to collect high-contrast BSE images of the agglutinates. Several images were collected to represent each single agglutinate. The contrast, brightness, and focus were adjusted to optimize each image collected. Histograms of the grayscale range for all images produced four 'peaks' corresponding to epoxy, glass, crystalline phases, and metal grains. We analyzed every grid point for 12 elements with a less than 1 micron electron beam using a CAMECA SX-50 electron probe microanalyzer (EPM). If any analysis was not within about 10% of the stoichiometry of a known lunar mineral, we considered that point to be nonmineralic. Our results show that there is a remarkable correspondence in the glass percentages obtained by the two methods. The EPM method may overestimate glass because secondary fluorescence from dusty clasts in agglutinitic glass can give the appearance of nonmineralic targets. The BSE method may underestimate glass because the diversity of compositions of agglutinitic glass may not be contained under one grayscale 'peak' during image analysis.

  10. Anatomy of individual agglutinates from a lunar highland soil

    NASA Astrophysics Data System (ADS)

    Basu, Abhijit; McKay, David S.; Morris, Richard V.; Wentworth, Susan J.

    1996-11-01

    We report results of our investigation of the relationship between values of Is/FeO (relative concentration of nanophase Fe0 divided by total FeO content), glass abundance, total Fe content, and degree of digestion of <20 μm clasts for 22 individual agglutinates (250-1000 μm) from the mature Apollo 16 soil 61181 (Is/FeO = 82 units in the <250 μm fraction). Agglutinates are important products of space weathering on the Moon, and they influence spectral observations at visible and near-IR wavelengths. Values of Is/FeO for individual agglutinates (250-1000 μm) within this single soil span a range from 3 to 262 units which is larger than the range observed for all Apollo 16 bulk soils (˜0 to 110 units). No correlation was observed between Is/FeO and glass abundance and FeO concentrations for either agglutinitic glass or whole agglutinate particles under investigation. Our results suggest that the variation in Is/FeO for agglutinates from a single soil may be in part a consequence of natural mixing processes on the Moon that produce highly-variable environments (with respect to surface exposure) for agglutinate formation and in part to variable kinetics of reactions in an agglutinate melt, which are influenced by a variety of factors including melt composition, temperature, impactor velocity, and quench rate. We cannot exclude but do not see evidence for other processes including addition of exotic agglutinates, micrometeoritic bombardment into compositionally-diverse microtargets, recycling of agglutinates, preferential melting of very fine soil particles, and production of nanophase Fe0 in amorphous rims of very fine irradiated lunar grains contributing to the observed variation of Is/FeO.

  11. Lectin-mediated agglutination of murine lymphoma cells. Cell surface deformability and reversibility of agglutination by saccharides.

    PubMed

    Nicolson, G L; Poste, G

    1979-07-01

    Agglutination of S49 mouse lymphoma cells by Ricinus communis I agglutinin can be reversed by the competing haptenic saccharide, lactose, soon after agglutination, but after further incubation in the absence of lectin the agglutination reaction could not be reversed by lactose and the cells remained as multicell aggregates. The irreversibility of S49 cell agglutination was time, temperature and lectin concentration dependent and its onset correlated with ultrastructurally observed deformation of adjacent cell surfaces and an increase in the proportion of adjacent cell surface areas in close apposition within multicell aggregates. Pretreatment of S49 cells with cytochalasin B or cytochalasin B plus vinblastine enhanced R. communis I agglutinin-mediated agglutination, while vinblastine alone and fluoride plus azide had essentially no effect. When drug-treated cells were agglutinated and then incubated in lectin-free drug-containing media for various times prior to lactose addition, the drug effects were more pronounced. Cytochalasin B alone or with vinblastine inhibited lactose reversal of S49 cell agglutination compared to the drug-free controls, while fluoride plus azide enhanced hapten reversibility. Electron microscopic analysis revealed that the onset of agglutination irreversibility correlated with cell surface deformation in the drug-treated cells. Cell aggregates that were more readily reversible by lactose (fluoride plus azide) were unchanged or less deformed, while S49 aggregates treated with cytochalasin B plus vinblastine were more deformed compared to controls without drugs. These experiments suggest a role for cell surface deformability as an important secondary effect during lectin-mediated cell agglutination of S49 lymphoma cells.

  12. Antibody blocks acquisition of bacterial colonization through agglutination.

    PubMed

    Roche, A M; Richard, A L; Rahkola, J T; Janoff, E N; Weiser, J N

    2015-01-01

    Invasive infection often begins with asymptomatic colonization of mucosal surfaces. A murine model of bacterial colonization with Streptococcus pneumoniae was used to study the mechanism for mucosal protection by immunoglobulin. In previously colonized immune mice, bacteria were rapidly sequestered within large aggregates in the nasal lumen. To further examine the role of bacterial agglutination in protection by specific antibodies, mice were passively immunized with immunoglobulin G (IgG) purified from antipneumococcal sera or pneumococcal type-specific monoclonal human IgA (hIgA1 or hIgA2). Systemically delivered IgG accessed the mucosal surface and blocked acquisition of colonization and transmission between littermates. Optimal protection by IgG was independent of Fc fragment and complement and, therefore, did not involve an opsonophagocytic mechanism. Enzymatic digestion or reduction of IgG before administration showed that protection required divalent binding that maintained its agglutinating effect. Divalent hIgA1 is cleaved by the pneumococcal member of a family of bacterial proteases that generate monovalent Fabα fragments. Thus, passive immunization with hIgA1 blocked colonization by an IgA1-protease-deficient mutant (agglutinated) but not the protease-producing wild-type parent (not agglutinated), whereas protease-resistant hIgA2 agglutinated and blocked colonization by both. Our findings highlight the importance of agglutinating antibodies in mucosal defense and reveal how successful pathogens evade this effect.

  13. Characterization of a rabbit cationic protein (CAP18) with lipopolysaccharide-inhibitory activity.

    PubMed Central

    Hirata, M; Shimomura, Y; Yoshida, M; Morgan, J G; Palings, I; Wilson, D; Yen, M H; Wright, S C; Larrick, J W

    1994-01-01

    Cationic antibacterial proteins (CAP) were purified from rabbit granulocytes, and the effects of CAP on lipopolysaccharide (LPS)-induced tissue factor generation by murine peritoneal macrophages and human blood monocytes were studied. CAP were purified from rabbit peritoneal leukocytes by using as an assay the agglutination of erythrocytes coated with Re-LPS. Two proteins with CAP activity, CAP18 (18 kDa) and CAP7 (7 kDa), were isolated by acid extraction, ethanol precipitation, affinity chromatography, gel filtration, and reverse-phase high-pressure liquid chromatography. On the basis of protein sequencing, CAP7 was identified as the C-terminal fragment of CAP18, designated CAP18(106-142). Various forms of LPS (S-LPS, Re-LPS, and lipid A) activate murine macrophages and human blood monocytes to generate tissue factor (tissue thromboplastin). Incubation of LPS for 18 h with partially purified CAP (heparin-Sepharose fraction) inhibited the capacity of LPS to induce tissue factor; however, purified CAP18 inhibited about 75% of the activity of S-LPS after 1 h of incubation. CAP more effectively inhibited S-LPS than Re-LPS or lipid A. Synthetic CAP18(106-142) inhibited LPS-induced tissue factor generation by murine macrophages. CAP18(106-142) has greater LPS-binding and LPS-neutralizing activities than CAP18. We hypothesize that CAP18 and the derivative peptide, CAP18(106-142), bind to LPS and alter the capacity of LPS to initiate disseminated intravascular coagulation. In this regard, CAP may have therapeutic potential for sepsis and endotoxin shock. Images PMID:8132348

  14. Erythrocyte membrane elasticity during in vivo ageing.

    PubMed

    Nash, G B; Wyard, S J

    1981-05-01

    Changes in the ability of senescent erythrocytes to pass through the microcirculation may cause them to be trapped in the spleen and removed from the blood. To help understand this process we have measured erythrocyte membrane elasticity, to see whether it changes during in vivo ageing. Human and rabbit red cells were fractionated by isopycnic sedimentation to obtain samples of aged and young cells. These were subjected to micropipette analysis in order to determine their membrane shear elastic modulus. We found that the membrane rigidity did not significantly alter as red cells aged. Previously we have also demonstrated that the changed size and shape of aged cells is unlikely to explain their removal from the circulation (Nash, G.B. and Wyard, S.J. (1981) Biorheology, in the press). Thus we conclude that the lifespan of erythrocytes is not determined by factors related to membrane flexibility or cell shape but may depend on changes in their viscous properties (as suggested by Williams, A.R. and Morris, D.R. (1980), Scand. J. Haematol. 24, 57--62).

  15. Identification of Babesia bigemina infected erythrocyte surface antigens containing epitopes conserved among strains.

    PubMed

    Shompole, S; McElwain, T F; Jasmer, D P; Hines, S A; Katende, J; Musoke, A J; Rurangirwa, F R; McGuire, T C

    1994-03-01

    The presence of previously uncharacterized antigens (new antigens) on the surface of intact erythrocytes infected with three strains of Babesia bigemina from Kenya and one each from Puerto Rico, Mexico, St. Croix, and Texcoco-Mexico was demonstrated by indirect immunofluorescent antibody (IFA) reactions. These antigens were not strain specific because antibodies in bovine immune serum to either the Mexico or Kenya isolates reacted with all seven strains tested. Homologous and heterologous immune serum antibodies bound a maximum of 83% and 55%, respectively, of intact erythrocytes infected with the Kenya-Ngong strain but not uninfected erythrocytes. Both sera caused agglutination of only infected erythrocytes. Antibodies eluted from the surface of glutaraldehyde (0.25%) fixed infected erythrocytes had IFA reaction patterns among strains similar to those of immune sera before elution. Eluted antibodies were used to determine if these antigens were protein and encoded by B. bigemina. Eluted antibodies bound seven parasite-encoded proteins of 240, 220, 66, 62, 58, 52 and 38 kDa in an erythrocyte surface-specific immunoprecipitation reaction of 35S-methionine labelled proteins. It was concluded that the surface of B. bigemina infected erythrocytes had parasite-encoded proteins and that these proteins had surface exposed epitopes that were conserved among the seven strains examined which were from two continents.

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

    SciTech Connect

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

    1986-03-05

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

  17. 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.

  18. Ultrastructural study of adhesion of enterotoxigenic Escherichia coli to erythrocytes and human intestinal epithelial cells.

    PubMed

    Knutton, S; Lloyd, D R; Candy, D C; McNeish, A S

    1984-05-01

    The adhesion to erythrocytes and human intestinal epithelial cells of enterotoxigenic Escherichia coli strains H10407, B2C, and H10407P, expressing colonization factor antigen I (CFA/I), CFA/II, and type 1 fimbriae, respectively, was examined by electron microscopy. CFA and type 1 fimbriae were visualized by negative staining in thin sections after en bloc staining with ruthenium red and by immune labeling with antisera raised against purified fimbriae. By negative and ruthenium red staining, CFA/I, CFA/II, and type 1 fimbriae were indistinguishable and appeared as approximately 7-nm-diameter hollow cylindrical structures up to 1.5 micron in length; strain B2C also produced 2- to 3-nm-diameter flexible fibrillar fimbriae. Bacteria producing CFA/I, CFA/II, and type 1 fimbriae adhered to and agglutinated human, bovine, and guinea pig erythrocytes, respectively; CFA/I and CFA/II also mediated attachment of bacteria to the brush border of isolated human duodenal enterocytes. Electron microscopy of agglutinated erythrocytes and enterocytes with adherent bacteria showed, in each case, that bacterial adhesion involved the formation of many interactions between the tips of fimbriae and receptors on the erythrocyte or enterocyte brush border membrane. Immune labeling allowed different fimbrial antigens mediating bacterial attachment to human enterocytes to be identified.

  19. 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. PMID:23746573

  20. 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.

  1. 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.

  2. Evolution of carbon isotopes, agglutinates, and the lunar regolith

    NASA Technical Reports Server (NTRS)

    Desmarais, D. J.; Basu, A.; Hayes, J. M.; Meinschein, W. G.

    1975-01-01

    Apollo 17 light-mantle soils and Apollo 15 Apennine Front soils are compared with respect to isotopic enrichment of C-13 and the maturity of the site. Analyses of soil-size fractions indicate that while the carbon concentration on particle surfaces remains relatively constant with increasing soil maturity, total surface-correlated carbon increases due to increasing total soil surface area. The role of agglutinates in the incorporation of surface-correlated carbon into aggregate grains is examined; agglutinates contain a major percentage of the carbon found in mature soil, and the volume-correlated carbon component in agglutinates apparently continues to increase after the surface-correlated carbon concentrations have reached a constant value. Constraints that may limit the carbon concentration in lunar soils to a value not greater than 300 micrograms/g are considered.

  3. 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.).

  4. Purification of Oncornaviruses by Agglutination with Concanavalin A

    PubMed Central

    Stewart, Margaret L.; Summers, Donald F.; Soeiro, Ruy; Fields, Bernard N.; Maizel, Jacob V.

    1973-01-01

    Concanavalin A (Con A) has been used to rapidly and selectively agglutinate murine and avian oncornavirions from culture medium or plasma. The agglutinated virus was concentrated rapidly and gently by low-speed centrifugation and solubilization with α-methyl mannoside. Infectious virus was purified 2.3 times with respect to nucleic-acid content, and more than 60% of its infectivity was recovered. Infectious particles of densities 1.18 and 1.16 g/cm3 were found in mouse cells infected with Friend virus. Con A reacted only with particles of density 1.16 g/cm3, indicating heterogeneity with respect to carbohydrate content or structure as well as buoyant density. Electron microscopy of virus agglutinated with Con A showed a zone of Con A-glycoprotein complexes averaging 12-15 nm in thickness. Images PMID:4514301

  5. 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.

  6. 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.

  7. Agglutinating serum for distinguishing Staphylococcus aureus of human biotype.

    PubMed

    Live, I

    1975-08-01

    Antiserum to Staphylococcus aureus strain 17 was treated with S. aureus strain 61218 until the antibodies against thermostable agglutinogen were removed. The absorbed serum agglutinated phage-typable as well as phageuntypable staphylococci of human biotype, whether recovered from people or from dogs. PMID:125241

  8. [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.

  9. STUDIES ON EXPERIMENTAL PLETHORA IN DOGS AND RABBITS

    PubMed Central

    Krumbhaar, Edward B.; Chanutin, Alfred

    1922-01-01

    1. The effects of repeated transfusions of blood on the blood-destroying and blood-forming apparatus of normal and splenectomized dogs and rabbits have been described. An anemia which developed despite continued blood transfusions in two dogs splenectomized during plethora has also been studied. 2. The decrease or absence of reticulocytes from the blood stream during plethora and their increase during "plethoric anemia" are evidently due to depression and activation of bone marrow activity. The response of the bone marrow is not immediate upon the onset of anemia, but is delayed for several days. 3. The blood volume studies have served to emphasize the constancy of plasma volume under extreme experimental conditions, and the adaptability of the circulatory system to large increases in total blood volume. 4. Blood destruction and elimination, as measured by urobilin excretion, are greatly increased during the stage of plethora, but still more so during "plethoric anemia." 5. Despite intravenous introductions of large quantities of nitrogen in the form of whole blood, the total nitrogen, urea, and ammonia in the urine and feces are not raised appreciably for some time after the onset of plethora. The normal organism is apparently able to store large quantities of blood or its decomposition products. Upon the onset of a "plethoric anemia," there is an increase in urinary total nitrogen and urea excretion, which was lowered during the course of the anemia. Albuminuria is also found at this time. Other nitrogenous constituents and phosphates show no striking changes. 6. Blood pigment, chiefly in the form of hemosiderin, is deposited in enormous quantities in the spleen, liver, lymph nodes, and bone marrow. It occurs chiefly in phagocytes, though in late stages large extracellular masses are found. Increased pigment deposition can still be found several months after transfusions have been stopped. 7. Phagocytes containing erythrocytes are found rarely, if at all, and

  10. Suppression of reagin synthesis in rabbits by passively administered antibody

    PubMed Central

    Strannegård, Ö.; Belin, L.

    1970-01-01

    The formation of rabbit antibodies, capable of sensitizing homologous skin, (reagins), was completely inhibited by passive administration of serum containing large quantities of 7S antibody 24 hours before or after antigen injection. No evident effect on reagin formation was noted when passive antibody was administered 8 days after antigen injection although some suppression of agglutinating antibody synthesis was observed. In rabbits not treated with passive antibody the injection of haemocyanin resulted in the formation of reagins reaching maximum serum concentrations 1 and 3 weeks following antigen injection. Both the `early' and `late' reagins persisted for a long time in the skin of injected rabbits, they appeared to have similar molecular size and both were devoid of PCA activity when injected into decomplemented rabbits. There was some indication that the `early' reagins may be more heat-labile than the `late' ones. A secondary reagin response was obtained in several animals which had shown a primary reagin response, but not in rabbits with inhibited primary response. The reagins formed in response to secondary antigen stimulation disappeared rapidly from the circulation, simultaneously with the rise in agglutinating antibody titres. The possible implications of the findings for the immunological treatment of allergic disorders is discussed. PMID:5420728

  11. Dielectric Properties and Ion Mobility in Erythrocytes

    PubMed Central

    Pauly, H.; Schwan, H. P.

    1966-01-01

    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. PMID:5970566

  12. Spreading of wheat germ agglutinin-induced erythrocyte contact by formation of spatially discrete contacts.

    PubMed

    Darmani, H; Coakley, W T; Hann, A C; Brain, A

    1990-06-01

    The time dependence of agglutination and cell-cell contact spreading in human erythrocytes exposed to wheat germ agglutinin (WGA) was characterized by light and electron microscopy. Cells (3 x 10(7)/mL) had a threshold lectin concentration in the range of 0.6-2.0 micrograms/mL for initial cell contact. Spreading was essentially completed within 60 and 2 min in undisturbed and gently agitated suspensions, respectively. The cells in large WGA agglutinates retained features of their initial disk form in contrast to the convex outlines of polycation or polyethylene glycol-induced agglutinates. Spreading of contact area was accompanied by development of a pattern of discrete contact regions separated by a distance of the order of 1 micron. Freeze fracture electron microscopy and studies with ferritin-labeled WGA showed no significant aggregation of intramembrane particles or specific lectin receptors under conditions when contact spreading occurred. It is argued that flow stress effects on cells in suspended agglutinates give rise to a situation where opposite membranes, at the leading edge of cell contact, are separated by a thin aqueous layer. When this intercellular water layer exceeds a critical length, it becomes unstable. The layer breaks up by surface wave development to form an array of intracellular water spaces. Formation of the aqueous spaces causes opposite membrane regions to move synchronously toward each other. Lectin molecules crosslink the wave crests to give spatially periodic contact points.

  13. 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.

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

    NASA Technical Reports Server (NTRS)

    Laul, J. C.; Smith, M. R.; Papike, J. J.; Simon, S. B.

    1984-01-01

    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.

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

    NASA Astrophysics Data System (ADS)

    Laul, J. C.; Smith, M. R.; Papike, J. J.; Simon, S. B.

    1984-11-01

    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.

  16. The Cell Adhesion Molecule “CAR” and Sialic Acid on Human Erythrocytes Influence Adenovirus In Vivo Biodistribution

    PubMed Central

    Wodrich, Harald; Billet, Olivier; Perreau, Matthieu; Hippert, Claire; Mennechet, Franck; Schoehn, Guy; Lortat-Jacob, Hugues; Dreja, Hanna; Ibanes, Sandy; Kalatzis, Vasiliki; Wang, Jennifer P.; Finberg, Robert W.; Cusack, Stephen; Kremer, Eric J.

    2009-01-01

    Although it has been known for 50 years that adenoviruses (Ads) interact with erythrocytes ex vivo, the molecular and structural basis for this interaction, which has been serendipitously exploited for diagnostic tests, is unknown. In this study, we characterized the interaction between erythrocytes and unrelated Ad serotypes, human 5 (HAd5) and 37 (HAd37), and canine 2 (CAV-2). While these serotypes agglutinate human erythrocytes, they use different receptors, have different tropisms and/or infect different species. Using molecular, biochemical, structural and transgenic animal-based analyses, we found that the primary erythrocyte interaction domain for HAd37 is its sialic acid binding site, while CAV-2 binding depends on at least three factors: electrostatic interactions, sialic acid binding and, unexpectedly, binding to the coxsackievirus and adenovirus receptor (CAR) on human erythrocytes. We show that the presence of CAR on erythrocytes leads to prolonged in vivo blood half-life and significantly reduced liver infection when a CAR-tropic Ad is injected intravenously. This study provides i) a molecular and structural rationale for Ad–erythrocyte interactions, ii) a basis to improve vector-mediated gene transfer and iii) a mechanism that may explain the biodistribution and pathogenic inconsistencies found between human and animal models. PMID:19119424

  17. STUDIES ON RABBIT LYMPHOCYTES IN VITRO

    PubMed Central

    Sell, Stewart; Gell, P. G. H.

    1965-01-01

    Rabbit lymphocytes may be stimulated in vitro with specific antiallotype sera to transform into "blast" cells and to synthesize DNA. This transformation only occurs when the donor cells are obtained from a rabbit having a given γ-globulin allotype (As4) and these cells are cultured in the presence of an antiserum prepared against the given allotype (As4). Heterologous (sheep, goat, and guinea pig) anti-rabbit γ-globulin sera also induce significant blast transformation and DNA synthesis in rabbit lymphocytes. Allotypic transformation and DNA synthesis are due to 7S antiallotype antibodies and do not require complement. The degree of transformation and rate of DNA synthesis is related to the concentration of antibody. Incubation of the appropriate cells with the antiallotype antibody for as short a time as 15 minutes results in a significant degree of "blast" transformation, indicating that the recognition of the antiallotype specificity in the cells and stimulation of the cellular changes leading to eventual transformation is rapid. The activity of the antiallotype sera as measured by transforming or haemagglutinating capacity, may be absorbed by lymphocytes of the appropriate allotype, but is not absorbed by lymphocytes from a donor rabbit not having the allotype to which the antiserum is directed. Transformation does not occur with mixtures of lymphocytes from different rabbits even if 1 donor is immunized against an allotype present in the other donor. Peripheral rabbit lymphocytes can also be induced to undergo "blast transformation" in vitro by phytohaemagglutinin and staphylococcal filtrate. The lack of demonstrable leucoagglutinins in staphylococcal filtrate and antiallotype serum indicates that agglutination is not a necessary prerequisite to the induction of blast transformation. PMID:14316952

  18. 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.

  19. 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

  20. 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

    2016-01-01

    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

  1. Fast Magnetic Field-Enhanced Linear Colloidal Agglutination Immunoassay.

    PubMed

    Daynès, Aurélien; Temurok, Nevzat; Gineys, Jean-Philippe; Cauet, Gilles; Nerin, Philippe; Baudry, Jean; Bibette, Jérôme

    2015-08-01

    We present the principle of a fast magnetic field enhanced colloidal agglutination assay, which is based on the acceleration of the recognition rate between ligands and receptors induced by magnetic forces. By applying a homogeneous magnetic field of 20 mT for only 7 s, we detect CRP (C-reactive protein) in human serum at a concentration as low as 1 pM for a total cycle time of about 1 min in a prototype analyzer. Such a short measurement time does not impair the performances of the assay when compared to longer experiments. The concentration range dynamic is shown to cover 3 orders of magnitude. An analytical model of agglutination is also successfully fitting our data obtained with a short magnetic pulse.

  2. Concanavalin A-induced agglutination of human leukemic and lymphoma cells.

    PubMed

    Maca, R D

    1976-04-01

    With a newly developed turbidometric method, concanavalin A was shown to agglutinate normal lymphocytes, lymphoma cells, and leukemic cells from chronic lymphocytic leukemia and from acute myelocytic and lymphocytic leukemia. However, there was a marked difference in the kinetics of this agglutination process. Leukemic blast cells and cells from a patient with convoluted lymphoma agglutinated poorly in this system. Conversely, the degree of agglutination for chronic lymphocytic leukemia cells was greater than that for the blast cells and also slightly greater than that for normal lymphocytes. Cultured cells from a Burkitt's lymphoma (Raji) and from a patient with poorly differentiated lymphoma agglutinated very rapidly with concanavalin A. Prior incubation of all cell types with neuraminidase markedly enhanced the agglutination process similar to that of trypsinization. Thus, these studies illustrate the usefulness of this method in quantitating the kinetics of agglutination of various human neoplastic cell types by concanavalin A. PMID:1063062

  3. Human erythrocytes have binding sites for beta-endorphin.

    PubMed Central

    Simpkins, C. O.; Chenet, B. P.; Kang, Y. H.; Mazorow, D. L.; Millar, D. B.; Hollis, V. W.

    1989-01-01

    Monoiodinated human beta-endorphin was found to bind specifically to human erythrocytes. Unlabeled beta-endorphin and beta-endorphin inhibited binding, but (-)naloxone, [D-Ala2, D-Leu5]-enkephalin, and leu- and met-enkephalin did not. Immunoelectron microscopy, using rabbit anti-beta-endorphin antibody, an antirabbit IgG secondary antibody, and complexed horseradish peroxidase, revealed that at low concentrations beta-endorphin binds to the cell surface. Electron spin resonance spectroscopy showed no effect of beta-endorphin on membrane fluidity. This receptor does not appear to conform to the characteristics of an opiate receptor. Images Figures 2 and 3 PMID:2560064

  4. Chlamydial hemagglutinin identified as lipopolysaccharide.

    PubMed

    Watkins, N G; Caldwell, H D; Hackstadt, T

    1987-08-01

    Chlamydial lipopolysaccharide (LPS) agglutinated mouse and rabbit erythrocytes but not human, guinea pig, or pronghorn antelope erythrocytes. Hemagglutination was not specific for Chlamydia spp., as rough LPSs from Coxiella burnetii and Escherichia coli also agglutinated erythrocytes from the same animal species. Nonagglutinated and agglutinated erythrocytes bound equivalent amounts of LPS, indicating that hemagglutination was not due to a specific interaction of chlamydial LPS with erythrocytes. Thus, hemagglutination by chlamydial LPS is not mediated by specific receptor-ligand interactions but is a property of the altered surface of the LPS-coated erythrocytes. PMID:3301820

  5. 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.

  6. Serum-Red Cell Interactions at Low Ionic Strength: Erythrocyte Complement Coating and Hemolysis of Paroxysmal Nocturnal Hemoglobinuria Cells*

    PubMed Central

    Jenkins, David E.; Hartmann, Robert C.; Kerns, Albert L.

    1967-01-01

    Complement coating and hemolysis were observed when erythrocytes from patients with paroxysmal nocturnal hemoglobinuria (PNH) were incubated in isotonic sucrose solution in the presence of small amounts of serum. Normal cells were likewise coated with complement components but did not hemolyze. Both normal and PNH erythrocytes reduced the hemolytic complement activity of the serum used in this reaction. Experience with other simple saccharides and related compounds suggests that the low ionic strength of the sucrose solution is the feature that permitted complement coating of red cells and hemolysis of PNH erythrocytes. Isotonic solutions of other sugars or sugar alcohols that do not readily enter human erythrocytes could be substituted for sucrose. The mechanism for these reactions may possibly relate to the agglutination observed with erythrocytes tested in the serum-sucrose system. Even though PNH hemolytic activity could be removed by prior heating of serum or barium sulfate treatment of plasma, the agglutination phenomenon still persisted. The in vitro conditions necessary for optimal sucrose hemolysis of PNH erythrocytes were described and compared with those of the classical acid hemolysis test. The requirement for less serum in the sucrose hemolysis system than needed in the standard acid hemolysis reaction makes certain experiments, especially those using large amounts of autologous PNH serum, much more feasible. Additional advantages of the sucrose hemolysis test are that it can be carried out at room temperature in the presence of oxalate and citrate and that critical pH control is not essential. To date, the sucrose hemolysis test has been a sensitive and specific one for PNH. A modified test used for screening purposes, the “sugar water” test, is very easy to perform. PMID:6025481

  7. Action of alpha-galactosidase from Clostridium sporogenes and coffee beans on blood group B antigen of erythrocytes. The effect on the viability of erythrocytes in circulation.

    PubMed

    Dybus, S; Aminoff, D

    1983-01-01

    The effect of alpha-galactosidase, purified from Clostridium sporogenes (Maebashi), was examined on erythrocytes from rats, rabbits and gibbons. The amount of galactose released by alpha-galactosidase from Cl. sporogenes and from coffee beans was compared. The amount of sialic acid released by Vibrio cholera sialidase was also determined. Loss of blood group B specificity following treatment with alpha-galactosidase was demonstrated with anti-B lectin. In animal models, removal of all the alpha-galactosyl residues with the coffee bean or clostridial alpha-galactosidase resulted in no change in the sequestration pattern of the treated erythrocytes over a period of several days. In contrast, erythrocytes treated with sialidase were rapidly sequestered from the circulation.

  8. 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.

  9. The significance of erythrocyte antigen site density

    PubMed Central

    Hoyer, Leon W.; Trabold, Norma C.

    1971-01-01

    The importance of antigen site density has been studied by means of a model passive hemolysis system using red cells coupled with sulfanilic acid groups. Relative site numbers were estimated from the covalent linkage of sulfanilic acid-35S to red cell membrane protein, and the effective antigen site number was determined with 125I-labeled rabbit IgG anti-sulfanilic acid (anti-S). Immune hemolysis was demonstrated for red cells which had greater than a threshold number of antigen sites, the value of which was different for normal human cells (80,000 sites/cell), cells from a patient with paroxysmal nocturnal hemoglobinuria (PNH) (40,000 sites/cell), and sheep red blood cells (RBC) (15,000 sites/cell). Cells with antigen site densities below these values did not hemolyze when tested with 1 mg/ml purified rabbit IgM anti-S. 2-8 times greater antigen site densities were required to obtain hemolysis with IgG anti-S. Above the threshold value, hemolysis titers were proportional to the antigen site number until maximal values were obtained. The greater hemolytic efficiency of IgM antibody was demonstrated in this system, and it was established that the magnitude of the difference was related to the test cell antigen site density. These data, taken with previously reported hemagglutination studies, have been used to develop a general classification of immune hemolysis and hemagglutination based on antigen site density and antibody class. It is suggested that the heterogeneity of blood group systems is caused by differences in the site separation of erythrocyte membrane antigens. PMID:5105661

  10. Occurrence of Toxoplasma gondii in domestic rabbits of Northeastern Brazil.

    PubMed

    de Lima, Débora Costa Viegas; Santos, André de Souza; da Silva, Luana Thamires Rapôso; de Melo, Renata Pimentel Bandeira; da Silva, José Givanildo; Júnior, José Wilton Pinheiro; Mota, Rinaldo Aparecido

    2016-09-01

    The present study aimed to conduct a serosurvey of toxoplasmosis in domestic rabbits of Northeastern Brazil. Blood samples and tissue fragments (brain, heart and diaphragm) were collected from 150 and 54 rabbits from the state of Pernambuco, Brazil, respectively. The serum samples were subjected to serological analysis (Modified Agglutination Test - MAT) and the tissue samples were assessed by PCR and histopathological analysis. Data collected through questionnaires were subjected to analysis of risk factors. According to the MAT and the PCR results, 6.7% (10/150; CI 3.2%-11.9%) of the rabbits were positive for anti-T. gondii antibodies and 9.25% (5/54) of the tissue fragments were positive for T. gondii DNA, respectively. Lesions associated with T. gondii infection, mainly characterized by granuloma, mononuclear cell infiltrates, degeneration areas and necrosis in brain and heart, were detected in the histopathological analysis. The risk factors associated with T. gondii infection identified in the present study were homemade food (odds ratio = 39.00) and contact between cats and rabbits (odds ratio = 52.00). This is the first report of toxoplasmosis in rabbits of Northeastern Brazil. The management problems identified in the present study must be corrected to reduce the frequency of positive animals in herds of rabbits. PMID:27447212

  11. 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

  12. 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

  13. Assessment of snapper (Pagrus auratus) natural IgM binding to bromelain treated sheep erythrocytes.

    PubMed

    Morrison, Richard N; Bruce Lyons, A; Nowak, Barbara F; Hayball, John D

    2005-01-01

    Normal snapper (Pagrus auratus Bloch and Schneider) serum was examined for natural IgM that binds to protease (bromelain) treated sheep erythrocytes (BrSRBC) in a model assay system that has been used to appraise natural IgM of various mammals. Normal snapper serum lysed BrSRBC while haemolysis was abrogated by heat inactivation of serum and in divalent cation-deficient conditions, indicative of classical complement mediated lysis. In addition, heat inactivated normal snapper serum agglutinated BrSRBC while phosphatidylcholine (PtC) liposomes partially inhibited both haemolysis and agglutination. Inhibition of haemolysis and agglutination may have been mediated by an interaction between immunoglobulin (Ig) and PtC as protein A purified snapper Ig bound to PtC liposomes. However it is not known if this binding was PtC specific nor if the binding was initiated by either the Fab and/or Fc domains of snapper Ig. BrSRBC plaque forming cells (PFC) were detected in the peritoneal cavity, spleen, head kidney and peripheral blood of normal snapper. The greatest proportion of BrSRBC PFC per B cell was within the peritoneal cavity followed by the spleen, peripheral blood and head kidney. Together, these data suggest that normal snapper serum may contain natural Ig that binds BrSRBC, activating the classical complement cascade.

  14. Effects of Phyllanthus sellowianus Müll Arg. extracts on the rheological properties of human erythrocytes.

    PubMed

    Buszniez, Patricia; Di Sapio, Osvaldo; Riquelme, Bibiana

    2014-11-01

    Phyllanthus sellowianus extracts have been used in Argentina since colonial times in the treatment of diabetes. The in vitro biorheological and hemoagglutinant action of different extracts of P. sellowianus bark on human erythrocytes (RBC) were studied. RBCs were incubated in vitro with four aqueous extracts: Maceration; Controlled Digestion (PD); Decoction; and Infusion. Biorheological parameters (deformability, membrane surface viscosity, elastic modulus, and dynamic viscolelasticity) were determined with an Erythrodeformeter, and erythrocyte adhesion was characterized by image digital analysis. Immunohematological assays in RBC incubated with all the extracts showed large globular aggregates and agglutination in human ABO blood group system. Isolated cell coefficient showed the increase of cell adhesion. Aggregated shape parameters were significantly higher than normal and they changed with the concentration, particularly of PD extracts. Rheological results showed that the extract biorheological action varies with the temperature used in the extract preparations. The results obtained are useful to study the action mechanism of extracts from P. sellowianus bark in order to evaluate its use as therapeutic agent in diabetes. Immunohematological Tests using ABO system showed its agglutinant power, which is of special interest in Immunohematology to be used as hemoclassifier.

  15. [Effect of erythrocyte preserved for different lengths of time on anti-D antibody identification with three blood matching tests].

    PubMed

    Xiao, Rui-Qing; Lin, Wu-Cun; Xu, Dan; Zeng, Jie; Wu, Jian-Jun; Zhao, Shu-Ming

    2003-10-01

    The specificity of the antigens and length of preservation time of erythrocytes are the interfering factors in blood group serological tests. In order to clarify the influence of preservation time of erythrocytes on the blood matching test, the titers of anti-D antibody were detected with papain method, BioVue cross matching card and DianaGel cross matching card in 7 series of panel red blood cells preserved for various length of time (0 to 9 months). The results showed that the titer of micro-column gel test (DianaGel card) was one tube higher than that of column agglutinating test (BioVue card). The titer of erythrocytes preserved for 9 months was as high as 256 tested by DianaGel card, but it was only 2 by papain method in the same anti-serum. It is suggested that there was no obvious difference between the results of micro-column gel test and column agglutinating test, and titer of papain method was the lowest. PMID:14575550

  16. Agglutinating secretory IgA preserves intestinal epithelial cell integrity during apical infection by Shigella flexneri.

    PubMed

    Mathias, Amandine; Longet, Stéphanie; Corthésy, Blaise

    2013-08-01

    Shigella flexneri, by invading intestinal epithelial cells (IECs) and inducing inflammatory responses of the colonic mucosa, causes bacillary dysentery. Although M cells overlying Peyer's patches are commonly considered the primary site of entry of S. flexneri, indirect evidence suggests that bacteria can also use IECs as a portal of entry to the lamina propria. Passive delivery of secretory IgA (SIgA), the major immunoglobulin secreted at mucosal surfaces, has been shown to protect rabbits from experimental shigellosis, but no information exists as to its molecular role in maintaining luminal epithelial integrity. We have established that the interaction of virulent S. flexneri with the apical pole of a model intestinal epithelium consisting of polarized Caco-2 cell monolayers resulted in the progressive disruption of the tight junction network and actin depolymerization, eventually resulting in cell death. The lipopolysaccharide (LPS)-specific agglutinating SIgAC5 monoclonal antibody (MAb), but not monomeric IgAC5 or IgGC20 MAbs of the same specificity, achieved protective functions through combined mechanisms, including limitation of the interaction between S. flexneri and epithelial cells, maintenance of the tight junction seal, preservation of the cell morphology, reduction of NF-κB nuclear translocation, and inhibition of proinflammatory mediator secretion. Our results add to the understanding of the function of SIgA-mediated immune exclusion by identifying a mode of action whereby the formation of immune complexes translates into maintenance of the integrity of epithelial cells lining the mucosa. This novel mechanism of protection mediated by SIgA is important to extend the arsenal of effective strategies to fight against S. flexneri mucosal invasion. PMID:23753631

  17. Agglutinating Secretory IgA Preserves Intestinal Epithelial Cell Integrity during Apical Infection by Shigella flexneri

    PubMed Central

    Mathias, Amandine; Longet, Stéphanie

    2013-01-01

    Shigella flexneri, by invading intestinal epithelial cells (IECs) and inducing inflammatory responses of the colonic mucosa, causes bacillary dysentery. Although M cells overlying Peyer's patches are commonly considered the primary site of entry of S. flexneri, indirect evidence suggests that bacteria can also use IECs as a portal of entry to the lamina propria. Passive delivery of secretory IgA (SIgA), the major immunoglobulin secreted at mucosal surfaces, has been shown to protect rabbits from experimental shigellosis, but no information exists as to its molecular role in maintaining luminal epithelial integrity. We have established that the interaction of virulent S. flexneri with the apical pole of a model intestinal epithelium consisting of polarized Caco-2 cell monolayers resulted in the progressive disruption of the tight junction network and actin depolymerization, eventually resulting in cell death. The lipopolysaccharide (LPS)-specific agglutinating SIgAC5 monoclonal antibody (MAb), but not monomeric IgAC5 or IgGC20 MAbs of the same specificity, achieved protective functions through combined mechanisms, including limitation of the interaction between S. flexneri and epithelial cells, maintenance of the tight junction seal, preservation of the cell morphology, reduction of NF-κB nuclear translocation, and inhibition of proinflammatory mediator secretion. Our results add to the understanding of the function of SIgA-mediated immune exclusion by identifying a mode of action whereby the formation of immune complexes translates into maintenance of the integrity of epithelial cells lining the mucosa. This novel mechanism of protection mediated by SIgA is important to extend the arsenal of effective strategies to fight against S. flexneri mucosal invasion. PMID:23753631

  18. A rapid slide agglutination test for the diagnosis of neurocysticercosis in the rural health set up

    PubMed Central

    Biswas, Rakhi; Parija, Subhash Chandra

    2011-01-01

    Background: Simple and rapid latex-based diagnostic tests have been used for detecting specific antigens or antibodies in several diseases. Aims: The aim of the present study was to standardize and evaluate the latex agglutination test (LAT) for the detection of Taenia solium metacestode antigen in the cerebrospinal fluid (CSF) and serum for the diagnosis of neurocysticercosis (NCC). Settings and Design: The study was conducted at Department of Microbiology, Jawaharlal Institute of Post graduate medical education and research after obtaining informed consent from the study subjects. Materials and Methods: In the present study, CSF and serum samples were collected from clinically suspected NCC, CT/MRI proven cases of NCC, non-cysticercal central nervous system infection control and from healthy control subjects. CSF was not collected from healthy controls. Polyclonal antisera raised in rabbits against porcine T. solium metacestode complete homogenate antigen, was used in the LAT to detect the antigen in the specimens. Statistical Analysis Used: The statistical analysis was carried out using Epi Info. The sensitivity, specificity, positive predictive value, and negative predictive value of the LAT were calculated. Results: The LAT exhibited sensitivity of 64.7% and specificity of 85.7% with CSF samples and sensitivity of 52.08% and specificity of 96% with serum samples. Conclusions: Results of the present study shows that the LAT can be employed as a moderately sensitive and specific test for the detection of T. solium metacestode antigen in the CSF and serum specimens for the diagnosis of NCC in poorly equipped laboratories. PMID:23508849

  19. 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

  20. An early Cambrian agglutinated tubular lophophorate with brachiopod characters

    NASA Astrophysics Data System (ADS)

    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.

    2014-05-01

    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.

  1. 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.

  2. Current status of erythrocyte substitutes.

    PubMed Central

    Biro, G. P.

    1983-01-01

    During the last two decades the search for alternatives to whole blood transfusions has led to promising developments in the field of erythrocyte substitutes. Hemoglobin solutions free of fragments of erythrocyte stroma and fluorocarbon emulsions are not blood-type-specific and appear likely to satisfy some proportion of our blood requirements. Both must be modified before becoming clinically useful. The oxygen affinity of the hemoglobin solution must be reduced and its intravascular persistence improved. Fluorocarbons cannot yet contribute significantly to the oxygen supply unless the patient breathes hyperbaric oxygen. Recent advances are leading to solutions for these problems. PMID:6344974

  3. 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…

  4. Isolation of a Factor from Apple that Agglutinates Erwinia amylovora12

    PubMed Central

    Romeiro, R.; Karr, A.; Goodman, R.

    1981-01-01

    Extracts prepared from apple seeds contain a factor (AF) capable of agglutinating cells of Erwinia amylovora. In drop agglutination tests, AF is more active in agglutinating an avirulent, acapsular strain of E. amylovora than a virulent, capsular strain. AF precipitates in agar plates with a receptor derived from boiled cells of avirulent acapsular strain and, therefore, can be located during fractionation by rocket electrophoresis. AF was heat-stable and had a pH optimum for agglutination near ≅3.6 pH. The agglutination activity was not affected by the presence of Mg2+, Ca2+, or EDTA. AF was separated into two fractions (AF I and AF II) by elution from a Bio-Gel P-100 column. The precipitin and agglutination activities associated with AF II were found to be present in a positively charged molecule which was sensitive to treatment with protease and trypsin and, hence, presumably resides in a protein. The approximate molecular weight of AF II was determined to be 12,600 daltons. Besides precipitating the receptor derived from cells of avirulent acapsular strain, AF II was capable of precipitating extracellular polysaccharide from cultures of virulent capsular strain, sodium polygalacturonate, and carboxymethylcellulose. These three polymers also inhibited the agglutination activity associated with AF II. AF II could be replaced by poly-l-lysines in both the precipitin and agglutination assays. In addition, in antigen absorption experiments, poly-l-lysines were found to remove the receptors for AF II from the boiled extracts of avirulent acapsular strain. Based on these observations, it is proposed that the activity of AF II resides in a highly positively charged protein which causes agglutination of bacterial cells by interacting on a charge-charge basis with negatively charged components on the surface of the bacterial cells. PMID:16661997

  5. Erythrocyte permeability to urea and water: comparative study in rodents, ruminants, carnivores, humans, and birds.

    PubMed

    Liu, Lifeng; Lei, Tianluo; Bankir, Lise; Zhao, Dan; Gai, Xiaodong; Zhao, Xuejian; Yang, Baoxue

    2011-01-01

    Mammalian erythrocytes exhibit high urea permeability (P (urea)) due to UT-B expression in their cytoplasmic membrane. This high P (urea) allows fast equilibration of urea in erythrocytes during their transit in the hyperosmotic renal medulla. It also allows more urea (in addition to that in plasma) to participate in counter-current exchange between ascending and descending vasa recta, thus improving the trapping of urea in the medulla and improving urine concentrating ability. To determine if P (urea) in erythrocytes is related to diet and urine concentrating ability, we measured P (urea) in erythrocytes from 11 different mammals and 5 birds using stopped-flow light scattering. Carnivores (dog, fox, cat) exhibited high P (urea) (in x10(-5) cm/s, 5.3 ± 0.6, 3.8 ± 0.5 and 2.8 ± 0.7, respectively). In contrast, herbivores (cow, donkey, sheep) showed much lower P (urea) (0.8 ± 0.2, 0.7 ± 0.2, 1.0 ± 0.1, respectively). Erythrocyte P (urea) in human (1.1 ± 0.2), and pig (1.5 ± 0.1), the two omnivores, was intermediate. Rodents and lagomorphs (mouse, rat, rabbit) had P (urea) intermediate between carnivores and omnivores (3.3 ± 0.4, 2.5 ± 0.3 and 2.4 ± 0.3, respectively). Birds that do not excrete urea and do not express UT-B in their erythrocytes had very low values (<0.1 × 10(-5) cm/s). In contrast to P (urea), water permeability, measured simultaneously, was relatively similar in all mammals. The species differences in erythrocytes P (urea) most probably reflect adaptation to the different types of diet and resulting different needs for concentrating urea in the urine.

  6. 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

  7. A lectin from the bark of the rubber tree (Hevea brasiliensis).

    PubMed

    Wititsuwannakul, R; Wititsuwannakul, D; Sakulborirug, C

    1998-01-01

    A protein isolated and purified from extracts of bark strips from mature rubber (Hevea brasilinsis) agglutinated erythrocytes from rabbits and all human blood types except AB. No agglutination was detected with erythrocytes from sheep, rates or mice. Proteolytic treatment of Hevea bark lectin (HBL) abolished haemagglutinin activity. The M(r) determined by SDS-PAGE was 40 k and that estimated from gel filtration was 140 k. Fetuin, asialofetuin, bovine submaxillary mucin and asialosubmaxillary mucin inhibited HBL-induced agglutination of rabbit erythrocytes. The HBL showed maximum haemagglutination activity over the pH range 4.5-9.5 and heat stability up to 60 degrees. PMID:9431671

  8. 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. PMID:25542240

  9. Commensal symbiosis between agglutinated polychaetes and sulfate-reducing bacteria.

    PubMed

    Guido, A; Mastandrea, A; Rosso, A; Sanfilippo, R; Tosti, F; Riding, R; Russo, F

    2014-05-01

    Pendant bioconstructions occur within submerged caves in the Plemmirio Marine Protected Area in SE Sicily, Italy. These rigid structures, here termed biostalactites, were synsedimentarily lithified by clotted-peloidal microbial carbonate that has a high bacterial lipid biomarker content with abundant compounds derived from sulfate-reducing bacteria. The main framework builders are polychaete serpulid worms, mainly Protula with subordinate Semivermilia and Josephella. These polychaetes have lamellar and/or fibrillar wall structure. In contrast, small agglutinated terebellid tubes, which are a minor component of the biostalactites, are discontinuous and irregular with a peloidal micritic microfabric. The peloids, formed by bacterial sulfate reduction, appear to have been utilized by terebellids to construct tubes in an environment where other particulate sediment is scarce. We suggest that the bacteria obtained food from the worms in the form of fecal material and/or from the decaying tissue of surrounding organisms and that the worms obtained peloidal micrite with which to construct their tubes, either as grains and/or as tube encompassing biofilm. Peloidal worm tubes have rarely been reported in the recent but closely resemble examples in the geological record that extend back at least to the early Carboniferous. This suggests a long-lived commensal relationship between some polychaete worms and heterotrophic, especially sulfate-reducing, bacteria.

  10. Hemolytic activity of ethylene glycol phenyl ether (EGPE) in rabbits.

    PubMed

    Breslin, W J; Phillips, J E; Lomax, L G; Bartels, M J; Dittenber, D A; Calhoun, L L; Miller, R R

    1991-10-01

    Studies were conducted to characterize the hemolytic effects of EGPE in rabbits following oral and dermal exposure, and to evaluate the in vitro hemolytic potential of EGPE and its major metabolite using rabbit red blood cells (RBC). Gavage administration of EGPE to female New Zealand White rabbits at 100, 300, 600, or 1000 mg/kg/day for up to 10 consecutive days (one dose/day) resulted in a dose-related intravascular hemolytic anemia. The hemolytic anemia was characterized by decreased RBC count, hemoglobin concentration, packed cell volume, hemoglobinuria, splenic congestion, renal tubule damage, and a regenerative erythroid response in the bone marrow. The hemolytic anemia was observed without alterations in RBC glutathione or methemoglobin. Phenoxyacetic acid (PAA) was identified as a major blood metabolite of EGPE. In vitro exposure of female rabbit erythrocytes indicated EGPE to be considerably more hemolytic than PAA. In a 90-day dermal study in which EGPE was applied to the skin of male and female New Zealand White rabbits 6 hr/day, 5 days/week, at doses up to 500 mg/kg/day, there was no indication of a hemolytic response. The only treatment-related effects were sporadic occurrences of slight erythema and scaling of skin at the site of test material application in high dose group male and female rabbits. However, erythema and scaling were not associated with gross or histopathologic changes and were not considered toxicologically significant.

  11. 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

  12. Hemolytic activity of plasma and urine from rabbits experimentally infected with Legionella pneumophila.

    PubMed

    Baine, W B; Rasheed, J K; Maca, H W; Kaufmann, A F

    1979-01-01

    Rabbits were infected with Legionella pneumophila by intravenous administration of allantoic fluid from eggs infected with this organism. Heated plasma from animals with severe illness caused by L. pneumophila lysed erythrocytes from guinea pigs in a radial hemolysis assay. Plasma from control rabbits did not lyse guinea pig erythrocytes in parallel assays. Urine from two of the infected animals also showed hemolytic activity. Attempts to induce illness in rabbits by intranasal administration of L. pneumohpila were less successful. Allantoic fluid from embrynated hen eggs developed hemolytic activity when maintained eithr in vitro at room temperature or in eggs whose embryos were killed by refrigeration. Hemolytic activity in filtrates of allantoic fluid from eggs infected with L. pneumophila, as previously reported, may not be due to the presence of bacterial hemolysins in the fluid. PMID:399383

  13. A comparative protein profile of mammalian erythrocyte membranes identified by mass spectrometry.

    PubMed

    Sharma, Savita; Punjabi, Vinny; Zingde, Surekha M; Gokhale, Sadashiv M

    2014-11-01

    A comparative analysis of erythrocyte membrane proteins of economically important animals, goat (Capra aegagrus hircus), buffalo (Bubalus bubalis), pig (Sus scrofa), cow (Bos tauras), and human (Homo sapiens) was performed. Solubilized erythrocyte membrane proteins were separated by sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE), visualized by staining the gels with Commassie Brilliant Blue (CBB), and identified by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Emerging results show that all major erythrocyte membrane proteins present in human are also seen in all the animals except for band 4.5 which could not be identified. Band 3 is seen as more intense and compact, band 4.1 appears as a doublet in all the animal erythrocyte membranes, band 4.2 exhibits a slightly higher molecular weight (Mr) in buffalo, and cow and band 4.9 has a higher Mr in all the animals relative to the human protein. In addition, there are two new bands in the goat membrane, band G1, identified as HSP 90α, and band G2 identified as HSP 70. A new band C2 identified as HSP 70 is also seen in cow membranes. Peroxiredoxin II is of lower intensity and/or higher Mr in the animals. The difference in size of the proteins possibly indicates the variations in the composition of the amino acids. The difference in intensity of the proteins among these mammalians highlights the presence of less or more number of copies of that protein per cell. This data complement the earlier observations of differences in the sialoglycoprotein profile and effect of proteases and neuraminidase on agglutination among the mammalian erythrocytes. This study provides a platform to understand the molecular architecture of the individual erythrocytes, and in turn the dependent disorders, their phylogenetic relationship and also generates a database of erythrocyte membrane proteins of mammals. The animals selected for this study are of economic importance as

  14. Evaluation of three commercial agglutination tests for the identification of Staphylococcus aureus.

    PubMed

    Pennell, D R; Rott-Petri, J A; Kurzynski, T A

    1984-10-01

    Three commercially available rapid slide agglutination tests for the identification of Staphylococcus aureus were evaluated with 354 recent clinical isolates (165 strains of S. aureus). The test results of two latex agglutination products, SeroSTAT Staph (Scott Laboratories, Inc.) and Staphylatex (American Micro Scan), and one hemagglutination product, Staphyloslide (BBL Microbiology Systems), were compared with the results of the tube coagulase test, which was read at 4 h (4-h tube coagulase test) and, if negative, again after overnight incubation at room temperature (24-h tube coagulase test). Discrepancies between agglutination and tube coagulase identifications were resolved by use of the thermonuclease, mannitol fermentation, and slide coagulase tests. All sensitivities, specificities, predictive values of a positive result, and predictive values of a negative result for the three agglutination tests were at least 98.8% and comparable with the 4-h tube coagulase test. Best results were obtained with the 24-h tube coagulase test, which yielded one false-negative and no false-positive tests. Agglutination identifications may be performed on organisms taken directly from a primary plate when sufficient growth is present. Kit agglutination procedures yield rapid and reliable identifications and are easy to perform. This study also demonstrates the usefulness of the 24-h tube coagulase test.

  15. Can erythrocytes release biologically active NO?

    PubMed

    Benz, Peter M; Fleming, Ingrid

    2016-01-01

    Under physiological conditions, endothelial cells and the endothelial nitric oxide (NO) synthase (eNOS) are the main source of NO in the cardiovascular system. However, several other cell types have also been implicated in the NO-dependent regulation of cell function, including erythrocytes. NO derived from red blood cells has been proposed to regulate erythrocyte membrane fluidity, inhibit platelet activation and induce vasodilation in hypoxic areas, but these proposals are highly controversial. In the current issue of Cell Communication and Signaling, an elegant study by Gambaryan et al., assayed NO production by erythrocytes by monitoring the activation of the platelet intracellular NO receptor, soluble guanylyl cyclase, and its downstream kinase protein kinase G. After systematically testing different combinations of erythrocyte/platelet suspensions, the authors found no evidence for platelet soluble guanylyl cyclase/protein kinase G activation by erythrocytes and conclude that erythrocytes do not release biologically active NO to inhibit platelet activation. PMID:27639852

  16. Induction of persistent autoimmune haemolytic anaemia in mice by combined use of rat erythrocyte preimmunization and chronic GVHR.

    PubMed Central

    Negoro, S; Takashima, T; Toba, H; Kishimoto, S

    1985-01-01

    Female F1 mice (BDF1: C57BL/6 x DBA/2, BCF1: C57BL/6 x C3H/HeN, BBF1: C57BL/6 x BALB/c) were preimmunized with rat erythrocytes and then received splenic T cells from parental female C57BL/6 mice injected with hydrocortisone acetate before killing. In BDF1 and BCF1 mice, direct Coombs' test (DCT) positivity persisted for more than 4 months in almost all mice. They also showed haematological sign of anaemia, and the survival of infused 51Cr-labelled parental C57BL/6 erythrocytes was reduced. In BBF1 mice, DCT positivity returned to negative within 2 months. Autoantibodies were eluted from DCT-positive erythrocytes and their specificities were examined. They were capable of binding equally well to erythrocytes from all strains of mice tested, including the donor mice. They also cross-reacted with rat erythrocytes but not with sheep, rabbit or human erythrocytes. Isotypes of the autoantibodies eluted were also examined. Interestingly, isotypes of the autoantibodies obtained from individual mice were confined to one or other subclass of IgG isotype: IgG1, IgG2a or IgG2b. PMID:3878325

  17. [The influence of extracorporeal laser radiation on structural indices of erythrocytes].

    PubMed

    Khetsuriani, R G; Aladashvili, L M; Arabuli, M B; Tophuria, D Z; Tchlikadze, N G

    2015-01-01

    Object of the research was to study the diffractometric indices of erythrocytes, while 1 ml of the blood of the experimental animals was irradiated extracorporally by helium-neon laser. For this purpose 1 ml blood was taken from normal weight, (1200 gr) grown up shinshila rabbits, that we divided into 7 groups and irradiated with 10 vat helium-neon laser during 0.5-1 minutes. After irradiation blood was injected back to the organism of rabbits. After 2-6 hours from irradiation blood was taken from veins, to study by electronic microscope and later to be processed by diffractometric analysis method. The examinations testify that after extracorporeal irradiation diffractometric characteristics of erythrocytes' membranes are lower than after general irradiation, which indicates to the different energetic possibilities of laser. The extracorporeal irradiation, performed by laser and input of radiated blood back to organism is considered to be a shock therapy from the side of erythrocytes, which promote the defense function of the organism itself. The base for the shock therapy should be important factors such as homeostasis, compensative-adaptive mechanisms and so on. Exactly this above mentioned should be manifested after the sensitized cells are led back to the body (1 ml of blood) and with their existence they should promote display of the defense mechanisms. PMID:25693223

  18. 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. PMID:25015439

  19. Optical tweezer for probing erythrocyte membrane deformability

    NASA Astrophysics Data System (ADS)

    Khan, Manas; Soni, Harsh; Sood, A. K.

    2009-12-01

    We report that the average rotation speed of optically trapped crenated erythrocytes is direct signature of their membrane deformability. When placed in hypertonic buffer, discocytic erythrocytes are subjected to crenation. The deformation of cells brings in chirality and asymmetry in shape that makes them rotate under the scattering force of a linearly polarized optical trap. A change in the deformability of the erythrocytes, due to any internal or environmental factor, affects the rotation speed of the trapped crenated cells. Here we show how the increment in erythrocyte membrane rigidity with adsorption of Ca++ ions can be exhibited through this approach.

  20. Infectious salmon anaemia virus replication and induction of alpha interferon in Atlantic salmon erythrocytes

    PubMed Central

    Workenhe, Samuel T; Kibenge, Molly JT; Wright, Glenda M; Wadowska, Dorota W; Groman, David B; Kibenge, Frederick SB

    2008-01-01

    Background Infectious salmon anaemia (ISA) virus (ISAV), which causes ISA in marine-farmed Atlantic salmon, is an orthomyxovirus belonging to the genus Isavirus, family Orthomyxoviridae. ISAV agglutinates erythrocytes of several fish species and it is generally accepted that the ISAV receptor destroying enzyme dissolves this haemagglutination except for Atlantic salmon erythrocytes. Recent work indicates that ISAV isolates that are able to elute from Atlantic salmon erythrocytes cause low mortality in challenge experiments using Atlantic salmon. Previous work on ISAV-induced haemagglutination using the highly pathogenic ISAV strain NBISA01 and the low pathogenic ISAV strain RPC/NB-04-0851, showed endocytosis of NBISA01 but not RPC/NB-04-0851. Real-time RT-PCR was used to assess the viral RNA levels in the ISAV-induced haemagglutination reaction samples, and we observed a slight increase in viral RNA transcripts by 36 hours in the haemagglutination reaction with NBISA01 virus when the experiment was terminated. However, a longer sampling interval was considered necessary to confirm ISAV replication in fish erythrocytes and to determine if the infected cells mounted any innate immune response. This study examined the possible ISAV replication and Type I interferon (IFN) system gene induction in Atlantic salmon erythrocytes following ISAV haemagglutination. Results Haemagglutination assays were performed using Atlantic salmon erythrocytes and one haemagglutination unit of the two ISAV strains, NBISA01 and RPC/NB-04-0851, of differing genotypes and pathogenicities. Haemagglutination induced by the highly pathogenic NBISA01 but not the low pathogenic RPC/NB-04-0851 resulted in productive infection as evidenced by increased ISAV segment 8 transcripts and increase in the median tissue culture infectious dose (TCID50) by 5 days of incubation. Moreover, reverse transcription (RT) quantitative PCR used to compare mRNA levels of key Type I IFN system genes in erythrocyte

  1. Defining the erythrocyte binding domains of Plasmodium vivax tryptophan rich antigen 33.5.

    PubMed

    Bora, Hema; Tyagi, Rupesh Kumar; Sharma, Yagya Dutta

    2013-01-01

    Tryptophan-rich antigens play important role in host-parasite interaction. One of the Plasmodium vivax tryptophan-rich antigens called PvTRAg33.5 had earlier been shown to be predominantly of alpha helical in nature with multidomain structure, induced immune responses in humans, binds to host erythrocytes, and its sequence is highly conserved in the parasite population. In the present study, we divided this protein into three different parts i.e. N-terminal (amino acid position 24-106), middle (amino acid position 107-192), and C-terminal region (amino acid position 185-275) and determined the erythrocyte binding activity of these fragments. This binding activity was retained by the middle and C-terminal fragments covering 107 to 275 amino acid region of the PvTRAg33.5 protein. Eight non-overlapping peptides covering this 107 to 275 amino acid region were then synthesized and tested for their erythrocyte binding activity to further define the binding domains. Only two peptides, peptide P4 (at 171-191 amino acid position) and peptide P8 (at 255-275 amino acid position), were found to contain the erythrocyte binding activity. Competition assay revealed that each peptide recognizes its own erythrocyte receptor. These two peptides were found to be located on two parallel helices at one end of the protein in the modelled structure and could be exposed on its surface to form a suitable site for protein-protein interaction. Natural antibodies present in the sera of the P. vivax exposed individuals or the polyclonal rabbit antibodies against this protein were able to inhibit the erythrocyte binding activity of PvTRAg33.5, its fragments, and these two synthetic peptides P4 and P8. Further studies on receptor-ligand interaction might lead to the development of the therapeutic reagent. PMID:23638151

  2. Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China.

    PubMed

    Meng, Qing-Feng; Wang, Wei-Lin; Ni, Xiao-Ting; Li, Hai-Bin; Yao, Gui-Zhe; Sun, Xiao-Lin; Wang, Wei-Li; Cong, Wei

    2015-12-01

    The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites. PMID:26797446

  3. Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China.

    PubMed

    Meng, Qing-Feng; Wang, Wei-Lin; Ni, Xiao-Ting; Li, Hai-Bin; Yao, Gui-Zhe; Sun, Xiao-Lin; Wang, Wei-Li; Cong, Wei

    2015-12-01

    The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.

  4. Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China

    PubMed Central

    Meng, Qing-Feng; Wang, Wei-Lin; Ni, Xiao-Ting; Li, Hai-Bin; Yao, Gui-Zhe; Sun, Xiao-Lin; Wang, Wei-Li; Cong, Wei

    2015-01-01

    The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites. PMID:26797446

  5. Prescription diets for rabbits.

    PubMed

    Proença, Laila Maftoum; Mayer, Jörg

    2014-09-01

    Dietary management can be used with drug therapy for the successful treatment of many diseases. Therapeutic nutrition is well-recognized in dogs and cats and is beginning to increase among other pet species, including rabbits. The nutritional component of some rabbit diseases (eg, urolithiasis) is not completely understood, and the clinician should evaluate the use of prescription diets based on the scientific literature and individual needs. Long-term feeding trials are needed to further evaluate the efficacy of prescription diets in rabbits. Prescription diets are available for selected diseases in rabbits, including diets for immediate-term, short-term, and long-term management. PMID:25155667

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

    PubMed

    Medina, Marjorie B; Shelver, Weilin L; Fratamico, Pina M; Fortis, Laurie; Tillman, Glenn; Narang, Neelam; Cray, William C; Esteban, Emilio; Debroy, Andchitrita

    2012-05-01

    Latex agglutination assays utilizing polyclonal antibodies were developed for the top six non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups. Rabbit antisera were affinity purified through protein A/G columns, and the isolated immunoglobulins (IgGs) were covalently immobilized onto polystyrene latex particles. The resulting latex-IgG complex had a protein (IgG) load of 0.20 to 0.28 mg/ml in a 1% latex suspension. Optimum conditions for the agglutination assay consisted of utilizing 20 μm l of latex-IgG reagent containing 2.0 to 2.8 μm g IgG in a 0.5% latex suspension. Agglutination or flocculation was observed almost instantly after mixing the colonies with the latex-IgG, indicating STEC strains. More than 100 target and nontarget strains were tested in more than 3,000 test replicates. All target organisms produced positive results, but three antisera (anti-O26, anti-O103, and anti-O145) cross-reacted with some other STECs. The anti-O103 and anti-O145 latex reagents cross-reacted with O26 strains, and the anti-O26 cross-reacted with O103 strains. The latex-IgG reagents are stable for at least 1 year and are easy to prepare. These agglutination assays can be used for identification of presumptive non-O157 STEC colonies from agar media. The techniques used to prepare the latex reagents also can be utilized for testing other STEC serogroups, other E. coli serotypes, or other pathogens to ensure safe foods to consumers.

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

    PubMed

    Medina, Marjorie B; Shelver, Weilin L; Fratamico, Pina M; Fortis, Laurie; Tillman, Glenn; Narang, Neelam; Cray, William C; Esteban, Emilio; Debroy, Andchitrita

    2012-05-01

    Latex agglutination assays utilizing polyclonal antibodies were developed for the top six non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups. Rabbit antisera were affinity purified through protein A/G columns, and the isolated immunoglobulins (IgGs) were covalently immobilized onto polystyrene latex particles. The resulting latex-IgG complex had a protein (IgG) load of 0.20 to 0.28 mg/ml in a 1% latex suspension. Optimum conditions for the agglutination assay consisted of utilizing 20 μm l of latex-IgG reagent containing 2.0 to 2.8 μm g IgG in a 0.5% latex suspension. Agglutination or flocculation was observed almost instantly after mixing the colonies with the latex-IgG, indicating STEC strains. More than 100 target and nontarget strains were tested in more than 3,000 test replicates. All target organisms produced positive results, but three antisera (anti-O26, anti-O103, and anti-O145) cross-reacted with some other STECs. The anti-O103 and anti-O145 latex reagents cross-reacted with O26 strains, and the anti-O26 cross-reacted with O103 strains. The latex-IgG reagents are stable for at least 1 year and are easy to prepare. These agglutination assays can be used for identification of presumptive non-O157 STEC colonies from agar media. The techniques used to prepare the latex reagents also can be utilized for testing other STEC serogroups, other E. coli serotypes, or other pathogens to ensure safe foods to consumers. PMID:22564929

  8. Induction of Suicidal Erythrocyte Death by Cantharidin

    PubMed Central

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

    2015-01-01

    The natural phosphoprotein phosphatase inhibitor cantharidin, primarily used for topical treatment of warts, has later been shown to trigger tumor cell apoptosis and is thus considered for the treatment of malignancy. Similar to apoptosis of tumor cells, erythrocytes may undergo eryptosis, a suicidal cell death characterized by cell shrinkage and translocation of cell membrane phosphatidylserine to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+-activity ([Ca2+]i), ceramide, oxidative stress and dysregulation of several kinases. Phosphatidylserine abundance at the erythrocyte surface was quantified utilizing annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ceramide from antibody binding, and reactive oxidant species (ROS) from 2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence. A 48 h treatment of human erythrocytes with cantharidin significantly increased the percentage of annexin-V-binding cells (≥10 μg/mL), significantly decreased forward scatter (≥25 μg/mL), significantly increased [Ca2+]i (≥25 μg/mL), but did not significantly modify ceramide abundance or ROS. The up-regulation of annexin-V-binding following cantharidin treatment was not significantly blunted by removal of extracellular Ca2+ but was abolished by kinase inhibitor staurosporine (1 μM) and slightly decreased by p38 inhibitor skepinone (2 μM). Exposure of erythrocytes to cantharidin triggers suicidal erythrocyte death with erythrocyte shrinkage and erythrocyte membrane scrambling, an effect sensitive to kinase inhibitors staurosporine and skepinone. PMID:26226001

  9. Induction of Suicidal Erythrocyte Death by Cantharidin.

    PubMed

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

    2015-08-01

    The natural phosphoprotein phosphatase inhibitor cantharidin, primarily used for topical treatment of warts, has later been shown to trigger tumor cell apoptosis and is thus considered for the treatment of malignancy. Similar to apoptosis of tumor cells, erythrocytes may undergo eryptosis, a suicidal cell death characterized by cell shrinkage and translocation of cell membrane phosphatidylserine to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+-activity ([Ca2+]i), ceramide, oxidative stress and dysregulation of several kinases. Phosphatidylserine abundance at the erythrocyte surface was quantified utilizing annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ceramide from antibody binding, and reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence. A 48 h treatment of human erythrocytes with cantharidin significantly increased the percentage of annexin-V-binding cells (≥10 mg/mL), significantly decreased forward scatter (≥25 mg/mL), significantly increased [Ca2+]i (≥25 mg/mL), but did not significantly modify ceramide abundance or ROS. The up-regulation of annexin-V-binding following cantharidin treatment was not significantly blunted by removal of extracellular Ca2+ but was abolished by kinase inhibitor staurosporine (1 mM) and slightly decreased by p38 inhibitor skepinone (2 mM). Exposure of erythrocytes to cantharidin triggers suicidal erythrocyte death with erythrocyte shrinkage and erythrocyte membrane scrambling, an effect sensitive to kinase inhibitors staurosporine and skepinone. PMID:26226001

  10. Screening of bacterial isolates for mannose-specific lectin activity by agglutination of yeasts.

    PubMed Central

    Mirelman, D; Altmann, G; Eshdat, Y

    1980-01-01

    A total of 393 clinical bacterial isolates were tested for their ability to agglutinate yeast cells of either Saccharomyces cerevisiae or Candida albicans. A positive agglutination of yeasts that could be prevented by methyl alpha-D-mannoside was taken as an indication for the possible presence of a mannose-specific lectin (carbohydrate-binding protein) on the surface of the tested bacteria. Agglutination tests on glass slides showed that 38% of all the isolates tested were positive in their capacity to agglutinate yeasts. Among the various strains tested, all isolates of Serratia marcescens, Proteus morganii, and Citrobacter diversus, as well as 94% of Klebsiella pneumoniae, were positive. On the other hand, only 46% of the Escherichia coli, 48% of the salmonellae, 44% of the Citrobacter freundii, and 71% of the Aeromonas hydrophila isolates were positive. A quantitative determination of the lectin activity done by observing the agglutination of yeasts in microtiter plates showed that S. marcescens isolates were the most avid binders to the yeast, whereas Klebsiella and Citrobacter isolates were the weakest. PMID:6989854

  11. Effect of Intramuscular or Intrahepatic Injections of Clostridium perfringens on Rabbit Serum Lactate Dehydrogenase 1

    PubMed Central

    Thomason, Dwayne

    1970-01-01

    Serum lactate dehydrogenase (LDH) activity, LDH isoenzyme pattern, phospholipase C activity, phosphorous level, hemoglobin, and erythrocyte osmotic fragility were followed in rabbits after intramuscular (IM) or intrahepatic (IH) injections of Clostridium perfringens. On the first day after IM injection, there was a drop in LDH activity; this was followed by an increase of LDH activity on the third and sixth day. On the seventh day, LDH activity began to decline, and by the ninth day it had almost returned to normal. On the sixth day after IM injection, there was an increase in serum LDH isoenzyme 5, hemoglobin, and erythrocyte osmotic fragility, but the increase of erythrocyte osmotic fragility and serum hemoglobin could not be attributed to phospholipase C activity since that enzyme was not detected nor was there an increase in serum phosphorus. C. perfringens was recovered by culturing the wound of IM-injected rabbits but not recovered from IH-injected rabbits. Rabbits injected IH showed no change from normal values in any of the tests performed. PMID:16557808

  12. Effects of a skullcap root supplement on haematology, serum parameters and antioxidant enzymes in rabbits on a high-cholesterol diet.

    PubMed

    Króliczewska, B; Miśta, D; Zawadzki, W; Wypchło, A; Króliczewski, J

    2011-02-01

    We studied the effects of Scutellaria baicalensis root on blood parameters and antioxidant enzyme activities in rabbits fed a high-cholesterol diet. Thirty-two New Zealand White rabbits were divided into four groups of eight animals each. They were fed a standard diet (C group), a diet with a 1% pure cholesterol supplement (CH group), a diet with a 1% pure cholesterol supplement and a 9% skullcap root supplement (CH+SR group), or a diet with a 9% skullcap root supplement (SR group). After 6 weeks, the rabbits fed the high-cholesterol diet had significantly decreased RBC and Hb levels and significantly increased MCV, MCH and Fe levels (p≤ 0.05). The skullcap root supplement had no adverse effects on the haematological parameters. The values for RBC, Hb, HCT, MCV, MCH and MCHC were similar in the skullcap root-treated and control rabbits. We also observed a remarkable elevation in the serum TC, LDL and TG levels at the end of the 6-week period. The rabbits fed the cholesterol diet showed decreased activity of the erythrocyte GSH-Px compared with the rabbits fed the basal diet. The GSH-Px activity was significantly higher in the rabbits fed the CH+SR diet than in those on the CH diet. The erythrocyte SOD activity was also significantly decreased in the rabbits on the CH diet. However, the CH+SR group rabbits showed significantly enhanced erythrocyte SOD activity. The SOD level in the CH+SR rabbits was 34.91 U/ml, which was a 23% increase (p≤ 0.05) in relation to the results for the CH group and only 15% diminished in relation to the control group. These results suggest that the dietary supplementation of skullcap root may improve rabbit antioxidant systems and protect against the risks from a high-cholesterol diet. PMID:20666864

  13. Erythrocyte membrane phosphatidylserine exposure in obesity.

    PubMed

    Solá, Eva; Vayá, Amparo; Martínez, Marcial; Moscardó, Antonio; Corella, Dolores; Santaolaria, Maria-Luisa; España, Francisco; Hernández-Mijares, Antonio

    2009-02-01

    It has been suggested that increased erythrocyte membrane phosphatidylserine (PS) exposure could contribute to hypercoagulability and hemorheological disturbances in obesity. The aim of our study was to evaluate PS exposure in obese patients and in a control group and to correlate this with hemorheological properties, i.e., erythrocyte aggregability (EA) and deformability, and to evaluate the effect of weight loss on these parameters. An anthropometric and analytical evaluation was performed at baseline and after 3 months on a diet (very low-calorie diet for 4 weeks and low-calorie diet for 2 months) on 49 severe or morbid obese patients (37 women, 12 men) and 55 healthy volunteers (39 women, 16 men). PS exposure on erythrocyte membrane was performed by flow cytometry. Erythrocyte aggregation was measured using the Myrenne MA(1) and the Sefam aggregometer. Erythrocyte deformability was determined in a stress diffractometer. Prothrombin fragment F1+2 (F1+2) was determined as a marker of the hypercoagulable state, and plasma malondialdehyde (MDA) as an indicator of oxidative stress. Obese patients had a higher EA index, higher PS exposure on erythrocyte membranes and higher levels of MDA and F1+2. The differences in erythrocyte aggregation and F1+2 between obese patients and the control group were maintained after adjusting for PS exposure. After 3 months of diet, a significant reduction in PS exposure on erythrocyte membrane was observed. Obese patients show increased PS exposure on erythrocyte membrane, with no effect on rheological properties. Increased PS exposure could contribute to hypercoagulability in these patients. Weight loss obtained with diet treatment reduces PS exposure on erythrocyte membrane.

  14. 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.

  15. Immunoglobulin determinants on the lymphocytes of normal rabbits

    PubMed Central

    Wolf, B.; Janeway, C. A.; Coombs, R. R. A.; Catty, D.; Gell, P. G. H.; Kelus, A. S.

    1971-01-01

    Experiments have been performed to see if `allelic exclusion' holds with regard to allotypic immunoglobulin determinants on circulating lymphocytes of the rabbit. The b locus allotypes As4 and As6 were investigated and these were measured by the mixed antiglobulin reaction. To investigate whether both allotypic determinants can be expressed on the membrane of any one lymphocyte, a lymphocyte suspension was treated with antibody reagents to both determinants. Adsorption of each antibody was shown by mixed agglutination with indicator red cells carrying either As4 or As6 immunoglobulin; one of the red cell suspensions was labelled with fluorescein isothiocyanate. When lymphocytes of adult heterozygous rabbits were tested, separate As4 and As6 lymphocytes were shown, but often more than 50 per cent of the lymphocytes exhibited both determinants on the same cell. Tests on an artificial mixture of homozygous As44 lymphocytes and homozygous As66 lymphocytes produced only distinct separate rosettes of the two types. Tests were performed on the lymphocytes of baby rabbits resulting from matings of As/46 does × homozygous As/44 bucks. At 2–3 weeks, when maternal As/46 immunoglobulin was present in easily detectable amounts in the sera of all baby rabbits, genetically As/44 animals could be clearly differentiated from genetically As/46 animals; correspondingly in matings with homozygous As/66 bucks. In litters of matings As/44 does × As/66 bucks, As6 determinants could be detected on the lymphocytes before the As6 immunoglobulin could be shown in the serum. In the very young (2–3 weeks) baby heterozygous As/46 rabbits, the majority of lymphocytes reacted as either As4 or As6 with a very much smaller percentage of mixed As46 cells. By the 12th week, the number of cells exhibiting both determinants had increased but not to so high a figure as found in the adults. PMID:4104284

  16. 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.

  17. Triggering of Programmed Erythrocyte Death by Alantolactone

    PubMed Central

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

    2014-01-01

    The sesquiterpene alantolactone counteracts malignancy, an effect at least in part due to stimulation of suicidal death or apoptosis of tumor cells. Signaling of alantolactone induced apoptosis involves altered gene expression and mitochondrial depolarization. Erythrocytes lack mitochondria and nuclei but may enter suicidal death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the erythrocyte surface. Cellular mechanisms involved in triggering of eryptosis include increase of cytosolic Ca2+-activity ([Ca2+]i) and oxidative stress. The present study explored, whether alantolactone stimulates eryptosis. To this end, erythrocyte volume was estimated from forward scatter, phosphatidylserine-exposure at the erythrocyte surface from FITC-annexin-V-binding, [Ca2+]i from Fluo3-fluorescence, ceramide abundance from binding of fluorescent antibodies, and oxidative stress from 2',7'-dichlorodihydrofluorescein-diacetate (DCFDA) fluorescence. As a result, a 48 h exposure of human erythrocytes to alantolactone (≥20 μM) significantly decreased erythrocyte forward scatter and increased the percentage of annexin-V-binding cells. Alantolactone significantly increased Fluo3 fluorescence (60 μM), ceramide abundance (60 μM) and DCFDA fluorescence (≥40 μM). The effect of alantolactone (60 μM) on annexin-V-binding was not significantly modified by removal of extracellular Ca2+. In conclusion, alantolactone stimulates suicidal erythrocyte death or eryptosis, an effect paralleled by increase of [Ca2+]i, ceramide abundance and oxidative stress. PMID:25533522

  18. Erythrocyte rouleau formation under polarized electromagnetic fields

    NASA Astrophysics Data System (ADS)

    Sebastián, José Luis; San Martín, Sagrario Muñoz; Sancho, Miguel; Miranda, José Miguel; Álvarez, Gabriel

    2005-09-01

    We study the influence of an external electromagnetic field of 1.8GHz in the formation or disaggregation of long rouleau of identical erythrocyte cells. In particular we calculate the variation of the transmembrane potential of an individual erythrocyte illuminated by the external field due to the presence of the neighboring erythrocytes in the rouleau, and compare the total electric energy of isolated cells with the total electric energy of the rouleau. We show that the polarization of the external electromagnetic field plays a fundamental role in the total energy variation of the cell system, and consequently in the formation or disaggregation of rouleau.

  19. 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.

  20. 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. PMID:26922300

  1. Comparison of two agglutination tests for differentiation between coagulase positive and coagulase negative staphylococci.

    PubMed

    Flesland, O

    1987-02-01

    Two agglutination tests, Monostaph (Bionor A/S, N-3700 Skien, Norway) and Staphaurex (Wellcome), for the identification of Staphylococcus aureus have been evaluated. Using the coagulase test as reference method both tests were equally reliable and in complete aggrement with the coagulase tube test when tested on 216 strains of Micrococcaceae.

  2. Development of a blocking latex agglutination test for the detection of antibodies to chicken anemia virus.

    PubMed

    Trinh, Dai Quang; Ogawa, Haruko; Bui, Vuong Nghia; Nguyen, Tham Thi Hong; Gronsang, Dulyatad; Baatartsogt, Tugsbaatar; Kizito, Mugimba Kahoza; AboElkhair, Mohammed; Yamaguchi, Shigeo; Nguyen, Viet Khong; Imai, Kunitoshi

    2015-09-01

    A blocking latex agglutination test (b-LAT) developed in this study was evaluated for the detection of antibodies against chicken anemia virus (CAV) in chickens. Polystyrene latex beads were coupled with a neutralizing monoclonal antibody (mAb) to CAV (mAb-beads). When mAb-beads were mixed with antigens prepared from the lysate of MDCC-MSB1 cells infected with CAV, agglutination occurred. A short pre-incubation of CAV antigens with CAV-specific antiserum inhibited the agglutination of mAb-beads. The test results were obtained within 5min. The specificity of b-LAT was evaluated using sera from specific pathogen-free chickens and sera containing antibodies to avian influenza virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus; nonspecific agglutination and cross-reactivity with antibodies to unrelated viruses were not observed. The examination of 94 serum samples collected from commercial breeder chickens of various ages (17-63 weeks) revealed good agreement (93.6%, Kappa value=0.82) between b-LAT and a virus neutralization test, known to be most sensitive and specific in the detection of antibodies to CAV. These results indicate that b-LAT, a simple and rapid test, is a useful and reliable tool in CAV serology.

  3. Agglutinating monoclonal antibodies that specifically recognize lipooligosaccharide A of Bordetella pertussis.

    PubMed Central

    Li, Z M; Cowell, J L; Brennan, M J; Burns, D L; Manclark, C R

    1988-01-01

    Monoclonal antibodies that specifically agglutinate strains of Bordetella pertussis having serotype 1 agglutinogen were uniquely reactive with the electrophoretically slow-migrating A form of lipooligosaccharide. These monoclonal antibodies should be useful for the structural analysis of B. pertussis lipooligosaccharide and for the establishment of a better-defined serogroup for Bordetella species. Images PMID:2893776

  4. Agglutinating monoclonal antibodies that specifically recognize lipooligosaccharide A of Bordetella pertussis.

    PubMed

    Li, Z M; Cowell, J L; Brennan, M J; Burns, D L; Manclark, C R

    1988-03-01

    Monoclonal antibodies that specifically agglutinate strains of Bordetella pertussis having serotype 1 agglutinogen were uniquely reactive with the electrophoretically slow-migrating A form of lipooligosaccharide. These monoclonal antibodies should be useful for the structural analysis of B. pertussis lipooligosaccharide and for the establishment of a better-defined serogroup for Bordetella species. PMID:2893776

  5. The specificity of antisera against Bordetella pertussis examined by bacterial agglutination.

    PubMed

    Fredriksen, J H; Frøholm, L O; Kjennerud, U

    1987-12-01

    The specificity of conventional, adsorbed antisera against agglutinogens 1, 2, and 3 of Bordetella pertussis was examined by slide agglutination and by agglutination in microtitre wells. Unadsorbed hyperimmune sera showed higher agglutinating activity against autologous or homologous cells than against cells of heterologous serotype. Adsorption of sera with heterologous cells increased the serotype specificity considerably. In spite of extensive adsorption, these anti-agglutinogen sera were still found to cross-react with B. parapertussis and/or B. bronchiseptica strains. Adsorption experiments with B. pertussis hyperimmune sera against serotype 1-, 1.2-, and 1.3-organisms demonstrated that the cross-reacting surface antigens differed from the agglutinogens 1, 2, and 3. Thus, in making species-specific reagents for diagnostic use it may be of value to include adsorption with B. parapertussis and probably with B. bronchiseptica. Limited data indicated that there is no need to use B. avium for adsorption. The agglutination assays were also used to test three monoclonal antibodies stated to be specific for the agglutinogens 1, 2, and 3, respectively. Some anomalous behaviour for the anti-agglutinogen 1 reagent was found, whereas the anti-agglutinogen 2 and 3 reagents corresponded well with the present polyclonal factor sera. PMID:2894108

  6. The incomplete anti-Rh antibody agglutination mechanism of trypsinized ORh+ red cells.

    PubMed Central

    Margni, R A; Leoni, J; Bazzurro, M

    1977-01-01

    The capacity for binding to trypsinized and non-trypsinized ORh+ red cells, of the IgG incomplete anti-Rh antibody and its F(ab')2 and Fc fragments has been investigated. An analysis has also been made of the capacity of non-specific human IgG, aggregated non-specific human IgG, human IgM (19S) and IgM (7S), and of fragments Fcgamma, Fcmu and Fc5mu to inhibit the agglutination of trypsinized ORh+ red cells by the IgG incomplete anti-Rh antibody. The results obtained indicate that these antibodies behave in a similar manner to that of nonprecipitating antibodies, and that the agglutination of trypsinized red cells seems to be a mixed reaction due to the interaction of an Fab fragment with its Rh antigenic determinant present in the surface of a red cell and the Fc of the same molecule with a receptor for Fc present in adjacent red cells. The trypsin treatment apparently results in the liberation of occult Fc receptors. It has also been demonstrated that in the agglutination of ORh+ red cells by IgG incomplete anti-Rh antibody in the presence of albumin, interaction must occur in some manner between the albumin and the Fc fragment since the F(ab')2 fragment does not give rise to agglutination under such conditions. Images Figure 1 PMID:415968

  7. 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…

  8. An integrated fiberoptic-microfluidic device for agglutination detection and blood typing.

    PubMed

    Ramasubramanian, Melur K; Alexander, Stewart P

    2009-02-01

    In this paper, an integrated fiberoptic-microfluidic device for the detection of agglutination for blood type cross-matching has been described. The device consists of a straight microfluidic channel through with a reacted RBC suspension is pumped with the help of a syringe pump. The flow intersects an optical path created by an emitter-received fiber optic pair integrated into the microfluidic device. A 650 nm laser diode is used as the light source and a silicon photodiode is used to detect the light intensity. The spacing between the tips of the two optic fibers can be adjusted. When fiber spacing is large and the concentration of the suspension is high, scattering phenomenon becomes the dominant mechanism for agglutination detection while at low concentrations and small spacing, optointerruption becomes the dominant mechanism. An agglutination strength factor (ASF) is calculated from the data. Studies with a variety of blood types indicate that the sensing method correctly identifies the agglutination reaction in all cases. A disposable integrated device can be designed for future implementation of the method for near-bedside pre-transfusion check. PMID:18815884

  9. 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

  10. Capsular gene typing of Streptococcus agalactiae compared to serotyping by latex agglutination.

    PubMed

    Yao, Kaihu; Poulsen, Knud; Maione, Domenico; Rinaudo, C Daniela; Baldassarri, Lucilla; Telford, John L; Sørensen, Uffe B Skov; Kilian, Mogens

    2013-02-01

    We evaluated three different PCR-based capsular gene typing methods applied to 312 human and bovine Streptococcus agalactiae (group B Streptococcus [GBS]) isolates and compared the results to serotyping results obtained by latex agglutination. Among 281 human isolates 27% could not be typed by latex agglutination. All 312 isolates except 5 could be typed by the three PCR methods combined. Two of these methods were multiplex assays. Among the isolates that were typeable by both latex agglutination and capsular gene typing, 94% showed agreement between the two methods. However, each of the PCR methods showed limitations. One of the methods did not include all 10 recognized serotypes, one misidentified eight isolates of serotypes Ib and IV as serotype Ia, and one did not distinguish between serotypes VII and IX. For five isolates that showed aberrant patterns in the capsular gene typing, long-range PCR targeting the cps operon disclosed large insertions or deletions affecting the cps gene cluster. A sensitive flow cytometric assay based on serotype-specific antibodies applied to 76 selected isolates that were nontypeable by latex agglutination revealed that approximately one-half of these did express capsular polysaccharide. A procedure for convenient and reliable capsular gene typing to be included in epidemiological and surveillance studies of S. agalactiae is proposed. PMID:23196363

  11. Development and evaluation of a latex agglutination test for the rapid serodiagnosis of tularemia.

    PubMed

    Rastawicki, Waldemar; Rokosz-Chudziak, Natalia; Chróst, Anna; Gierczyński, Rafał

    2015-05-01

    A latex agglutination test (LAT) was developed for a rapid detection of antibodies against Francisella tularensis. The assay is performed by mixing serum with antigen-coated latex beads and read within 5 min. Developed LAT has been proved to be a specific, sensitive, fast, easy-to-perform and cost-efficient tool for the routine diagnosis of tularemia.

  12. The Molecular Basis of Erythrocyte Shape

    NASA Astrophysics Data System (ADS)

    Elgsaeter, Arnljot; Stokke, Bjorn T.; Mikkelsen, Arne; Branton, Daniel

    1986-12-01

    Recent discoveries about the molecular organization and physical properties of the mammalian erythrocyte membrane and its associated structural proteins can now be used to explain, and may eventually be used to predict, the shape of the erythrocyte. Such explanations are possible because the relatively few structural proteins of the erythrocyte are regularly distributed over the entire cytoplasmic surface of the cell membrane and because the well-understood topological associations of these proteins seem to be stable in comparison with the time required for the cell to change shape. These simplifications make the erythrocyte the first nonmuscle cell for which it will be possible to extend our knowledge of molecular interactions to the next hierarchical level of organization that deals with shape and shape transformations.

  13. The Erythrocyte Ghost Is a Perfect Osmometer

    PubMed Central

    Kwant, W. O.; Seeman, Philip

    1970-01-01

    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°. 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. PMID:5413078

  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. 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

  16. 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.

  17. 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. PMID:25769039

  18. A critical role for the regulation of Syk from agglutination to aggregation in human platelets.

    PubMed

    Shih, Chun-Ho; Chiang, Tin-Bin; Wang, Wen-Jeng

    2014-01-10

    Agglucetin, a tetrameric glycoprotein (GP) Ibα agonist from Formosan Agkistrodon acutus venom, has been characterized as an agglutination inducer in human washed platelets (WPs). In platelet-rich plasma (PRP), agglucetin dramatically elicits a biphasic response of agglutination and subsequent aggregation. For clarifying the intracellular signaling events from agglutination to aggregation in human platelets, we examined the essential signaling molecules involved through the detection of protein tyrosine phosphorylation (PTP). In WPs, an anti-GPIbα monoclonal antibody (mAb) AP1, but not a Src kinase inhibitor PP1, completely inhibited agglucetin-induced agglutination. However, PP1 but not AP1 had a potent suppression on platelet aggregation by a GPVI activator convulxin. The PTP analyses showed agglucetin alone can cause a weak pattern involving sequential phosphorylation of Lyn/Fyn, Syk, SLP-76 and phospholipase Cγ2 (PLCγ2). Furthermore, a Syk-selective kinase inhibitor, piceatannol, significantly suppressed the aggregating response in agglucetin-activated PRP. Analyzed by flow cytometry, the binding capacity of fluorophore-conjugated PAC-1, a mAb recognizing activated integrin αIIbβ3, was shown to increase in agglucetin-stimulated platelets. Again, piceatannol but not PP1 had a concentration-dependent suppression on agglucetin-induced αIIbβ3 exposure. Moreover, the formation of signalosome, including Syk, SLP-76, VAV, adhesion and degranulation promoting adapter protein (ADAP) and PLCγ2, are required for platelet aggregation in agglucetin/fibrinogen-activated platelets. In addition, GPIbα-ligation via agglucetin can substantially promote the interactions between αIIbβ3 and fibrinogen. Therefore, the signal pathway of Lyn/Fyn/Syk/SLP-76/ADAP/VAV/PLCγ2/PKC is sufficient to trigger platelet aggregation in agglucetin/fibrinogen-pretreated platelets. Importantly, Syk may function as a major regulator for the response from GPIbα-initiated agglutination to

  19. A critical role for the regulation of Syk from agglutination to aggregation in human platelets.

    PubMed

    Shih, Chun-Ho; Chiang, Tin-Bin; Wang, Wen-Jeng

    2014-01-10

    Agglucetin, a tetrameric glycoprotein (GP) Ibα agonist from Formosan Agkistrodon acutus venom, has been characterized as an agglutination inducer in human washed platelets (WPs). In platelet-rich plasma (PRP), agglucetin dramatically elicits a biphasic response of agglutination and subsequent aggregation. For clarifying the intracellular signaling events from agglutination to aggregation in human platelets, we examined the essential signaling molecules involved through the detection of protein tyrosine phosphorylation (PTP). In WPs, an anti-GPIbα monoclonal antibody (mAb) AP1, but not a Src kinase inhibitor PP1, completely inhibited agglucetin-induced agglutination. However, PP1 but not AP1 had a potent suppression on platelet aggregation by a GPVI activator convulxin. The PTP analyses showed agglucetin alone can cause a weak pattern involving sequential phosphorylation of Lyn/Fyn, Syk, SLP-76 and phospholipase Cγ2 (PLCγ2). Furthermore, a Syk-selective kinase inhibitor, piceatannol, significantly suppressed the aggregating response in agglucetin-activated PRP. Analyzed by flow cytometry, the binding capacity of fluorophore-conjugated PAC-1, a mAb recognizing activated integrin αIIbβ3, was shown to increase in agglucetin-stimulated platelets. Again, piceatannol but not PP1 had a concentration-dependent suppression on agglucetin-induced αIIbβ3 exposure. Moreover, the formation of signalosome, including Syk, SLP-76, VAV, adhesion and degranulation promoting adapter protein (ADAP) and PLCγ2, are required for platelet aggregation in agglucetin/fibrinogen-activated platelets. In addition, GPIbα-ligation via agglucetin can substantially promote the interactions between αIIbβ3 and fibrinogen. Therefore, the signal pathway of Lyn/Fyn/Syk/SLP-76/ADAP/VAV/PLCγ2/PKC is sufficient to trigger platelet aggregation in agglucetin/fibrinogen-pretreated platelets. Importantly, Syk may function as a major regulator for the response from GPIbα-initiated agglutination to

  20. 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

  1. Triggering of Erythrocyte Death by Triparanol

    PubMed Central

    Officioso, Arbace; Manna, Caterina; Alzoubi, Kousi; Lang, Florian

    2015-01-01

    The cholesterol synthesis inhibitor Triparanol has been shown to trigger apoptosis in several malignancies. Similar to the apoptosis of nucleated cells, erythrocytes may enter eryptosis, the suicidal death characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include oxidative stress which may activate erythrocytic Ca2+ permeable unselective cation channels with subsequent Ca2+ entry and increase of cytosolic Ca2+ activity ([Ca2+]i). The present study explored whether and how Triparanol induces eryptosis. To this end, phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, hemolysis from hemoglobin release, [Ca2+]i from Fluo3-fluorescence, and ROS formation from 2’,7’-dichlorodihydrofluorescein diacetate (DCFDA) dependent fluorescence. As a result, a 48 h exposure of human erythrocytes to Triparanol (20 µM) significantly increased DCFDA fluorescence and significantly increased Fluo3-fluorescence. Triparanol (15 µM) significantly increased the percentage of annexin-V-binding cells, and significantly decreased the forward scatter. The effect of Triparanol on annexin-V-binding was significantly blunted, but not abolished by removal of extracellular Ca2+. In conclusion, Triparanol leads to eryptosis, the suicidal erythrocyte death characterized by cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane. Triparanol is at least in part effective by stimulating ROS formation and Ca2+ entry. PMID:26305256

  2. [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.

  3. The hematologic effects of low intensity 650 nm laser irradiation on hypercholesterolemia rabbits

    PubMed Central

    Wang, Hong; Deng, Juan; Tu, Wenjun; Zhang, Lin; Chen, Hongli; Wu, Xiaoguang; Li, Yingxin; Sha, Hong

    2016-01-01

    Purpose: To test the hematologic effects of low intensity 650 nm laser irradiation on hypercholesterolemia rabbits. Methods: Ten male big-eared rabbits were selected from hypercholesterolemia animal model-making and divided into model group and laser treatment groups. Five normal rabbits were selected as control group. Auricle root irradiation of 650 nm laser 100 mW in 30 min were applied on treatment group twice a day, treatment of 6 days in one week, 20 weeks course of treatment. Changes in blood lipid, microcirculation, rheological properties, and aggregation morphology of erythrocytes were observed every two weeks. Histopathological examination were performed in the end of experiments. Results: After 20 weeks’ treatment, triglyceride (TG), cholesterol (CHO), high density lipoprotein (HDL) and low density lipoprotein (LDL) of serum in hypercholesterolemia groups showed less changes in the first 4 weeks, butdifferent decreasing trends were shownin the next 16 weeks’ therapy. Erythrocytes aggregation of model groups showed rouleau state, while red blood cells in control group showed fine homodisperse, erythrocytes in treatment group performed better dispersion than model groups. Erythrocyte deformation index (DI) and blood flow value showed a statistic improvement in treatment group than control and model group (P<0.01). Value of DI in treatment group decreased after 18 weeks than values before therapy (P<0.01). Varying degrees of Aorta plaque formation was observed for individual difference in model and treatment groups, while no plaque was found in control group. Conclusions: Low energy laser improve microcirculation, rheological properties and blood lipid that might be related with erythrocytes aggregation and deformability. PMID:27347336

  4. 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).

  5. Low lead doses and atherogenic diet in rabbits: biochemical results in blood.

    PubMed

    Speich, M; Métayer, C; Arnaud, P; Nguyen, V G; Bousquet, B; Boiteau, H L

    1983-01-01

    Changes in concentrations of blood lead, cholesterol, triacylglycerols, GOT and GPT enzymes, phosphorus, proteins, plasma calcium and magnesium, and erythrocyte magnesium were studied for 25 weeks in 4 groups of 6 rabbits each receiving different diets: I (controls), II (water with 9.66 mumol/l of lead), III (atherogenic) and IV (atherogenic + 9.66 mumol/l of lead). Differences observed, with respect to the diets, for lead, cholesterol, enzymes, proteins and erythrocyte magnesium were evident from the 42nd day on. Low lead doses raised the levels of cholesterol and triacylglycerols in group II, whereas triacylglycerols were lower in the plasma of group IV in comparison with group III and were accumulated in the aorta. The elevation of enzymes in groups III and IV indicated heart and liver dysfunctions. There was a significant drop in erythrocyte magnesium in groups III and IV, which was especially marked in the latter group, with an interaction between the two diets.

  6. A microtitration agglutination test for detecting group E streptococcus infection in swine.

    PubMed

    Armstrong, C H; Wood, R L; Wessman, G E

    1982-04-01

    A microtitration agglutination test was developed and evaluated for detecting infection of swine with group E streptococci type IV, the most common causative agent of streptococcic lymphadenitis of swine. Whole cell agglutinogens representing group and type antigens of group E streptococci were tested in the microtitration agglutination test against reference antisera to Streptococcus groups A, B, C, D, E, F, G. H, K, L, M, N, O, P, Q, R, S and U, as well as specific antisera to types II, IV and V of group E. Group E specific agglutinogens were unsatisfactory in the microtitration agglutination test because of cross reactions with group P and U antisera and because of poor reproducibility of the test. Type specific agglutinogens of group E streptococci reacted only with their respective homologous antisera and not with any heterologous group antisera. None of the group E streptococci agglutinogens reacted with 52 normal swine sera. Agglutinogen made from group E streptococci type IV was selected for further evaluation in the microtitration agglutination test because group E streptococci types II and V are considered to be of minor importance in the etiology of streptococcic lymphadenitis of swine. Swine experimentally infected with a type IV strain developed significant titers in the microtitration agglutination test. All swine tested negative before exposure and seroconverted (titer >/=4) two to six weeks postexposure.The microtitration agglutination test was used by two different laboratories to test 187 duplicate samples of serum from infected swine. A total of 94.1% of the tests were read at either the same titer (48.1%) or a difference of not more than one dilution (46.0%) at the two laboratories. There was disagreement between the two laboratories in the test-positive test-negative status of 19 of the sera (10.2%). Titers of two of the sera differed by two dilutions (<4 at one laboratory and 8 at the other). The remaining 17 sera differed in titer by only one

  7. 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.

  8. Evaluation of a latex agglutination assay for the identification of Burkholderia pseudomallei and Burkholderia mallei.

    PubMed

    Duval, Brea D; Elrod, Mindy G; Gee, Jay E; Chantratita, Narisara; Tandhavanant, Sarunporn; Limmathurotsakul, Direk; Hoffmaster, Alex R

    2014-06-01

    Cases of melioidosis and glanders are rare in the United States, but the etiologic agents of each disease (Burkholderia pseudomallei and Burkholderia mallei, respectively) are classified as Tier 1 select agents because of concerns about their potential use as bioterrorism agents. A rapid, highly sensitive, and portable assay for clinical laboratories and field use is required. Our laboratory has further evaluated a latex agglutination assay for its ability to identify B. pseudomallei and B. mallei isolates. This assay uses a monoclonal antibody that specifically recognizes the capsular polysaccharide produced by B. pseudomallei and B. mallei, but is absent in closely related Burkholderia species. A total of 110 B. pseudomallei and B. mallei were tested, and 36 closely related Burkholderia species. The latex agglutination assay was positive for 109 of 110 (99.1% sensitivity) B. pseudomallei and B. mallei isolates tested.

  9. 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.

  10. Widal agglutination test − 100 years later: still plagued by controversy

    PubMed Central

    Olopoenia, L.; King, A.

    2000-01-01

    We review the significance of the Widal agglutination test in the diagnosis of typhoid fever. Over 100 years since its introduction as a serologic means of detecting the presence of typhoid fever, the Widal test continues to be plagued with controversies involving the quality of the antigens used and interpretation of the result, particularly in endemic areas. Areas of concern with clinical and laboratory significance discussed in this review include: the techniques of test performance, interpretation of results, limitation of the value of the test results in endemic typhoid areas, the quality of the antigens used, and alternative diagnostic tests.


Keywords: Widal agglutination test; typhoid fever PMID:10644383

  11. [Use of a latex-agglutination test for determining anti-measles antibody titer].

    PubMed

    Aleksander, S K; Lukin, Iu V; Iuminova, N V; Krasnova, V P; Andzhaparidze, O G

    1995-01-01

    A new diagnostic agent for microtitration of antimeasles antibodies, making use of polyacrolein microspheres conjugated with purified measles virus has been developed. Parallel titration of blood sera of children and adults in latex agglutination test and in routine test (hemagglutination inhibition, passive hemagglutination, immunofluorescent tests) demonstrated a sufficient specificity of the new test, sensitivity compatible to that of hemagglutination inhibition, and correlation of the results of all tests.

  12. Blood viscosity: influence of erythrocyte deformation.

    PubMed

    Chien, S; Usami, S; Dellenback, R J; Gregersen, M I

    1967-08-18

    Suspensions of canine and human erythocytes hardened with acetaldehyde differ from the suspensions of normal erythrocytes with respect to their rheological behavior. Normal erythrocytes can be packed by centrifugation so that the sediment volume is nearly 100 percent cells, but the hardened erythrocytes (RBC's) can be packed only to approximately 60 percent cells. At the same cell percentage the viscosity of the hardened RBC suspension is higher than that of the suspension of normal erythocytes. An increase in shear stress deforms the normal erythocytes and lowers the suspension viscosity, but has no influence on the viscosity of the hardened cell suspension. In blood with high cell percentages, the shear deformation of normal RBC's plays an important role in reducing viscosity and facilitating flow at high shear stresses. PMID:17842793

  13. Comparison of type 2 and type 6 fimbriae of Bordetella pertussis by using agglutinating monoclonal antibodies.

    PubMed Central

    Li, Z M; Brennan, M J; David, J L; Carter, P H; Cowell, J L; Manclark, C R

    1988-01-01

    Two types of fimbriae have been identified on the pathogenic gram-negative organism Bordetella pertussis. Monoclonal antibodies to these fimbriae were produced to better understand the role of fimbriae as serotype-specific agglutinogens and to investigate the antigenic relationship between these fimbriae. Three monoclonal antibodies were identified that specifically agglutinated B. pertussis cells containing the U.S. Reference Factor 2 agglutinogen, and six monoclonal antibodies were produced that agglutinated only those strains containing the U.S. Reference Factor 6 agglutinogen. Indirect immunofluorescence studies and immunogold electron microscopy demonstrated that these monoclonal antibodies bind to an outer membrane component on serotype-specific strains of B. pertussis. All of the monoclonal antibodies reacted with native or partially assembled type-specific fimbriae but not with monomeric fimbrial subunits as indicated by Western blot (immunoblot) analysis. The fimbrial agglutinogens recognized by the monoclonal antibodies were also uniquely reactive with either U.S. Reference Factor 2 or 6 antiserum (Eldering agglutinogen 2 or 6 polyclonal antiserum) in an indirect ELISA. No cross-reactivity of the monoclonal antibodies with the unrelated fimbriae was observed in any of the comparative immunological studies. Some of the monoclonal antibodies agglutinated certain strains of B. bronchiseptica, suggesting that this closely related species can contain antigenically similar fimbriae. These monoclonal antibodies should prove useful for further structural and functional analysis of Bordetella fimbriae and for studies on the role that these antigens play in prevention of infection and disease. Images PMID:2903125

  14. Comparison of type 2 and type 6 fimbriae of Bordetella pertussis by using agglutinating monoclonal antibodies.

    PubMed

    Li, Z M; Brennan, M J; David, J L; Carter, P H; Cowell, J L; Manclark, C R

    1988-12-01

    Two types of fimbriae have been identified on the pathogenic gram-negative organism Bordetella pertussis. Monoclonal antibodies to these fimbriae were produced to better understand the role of fimbriae as serotype-specific agglutinogens and to investigate the antigenic relationship between these fimbriae. Three monoclonal antibodies were identified that specifically agglutinated B. pertussis cells containing the U.S. Reference Factor 2 agglutinogen, and six monoclonal antibodies were produced that agglutinated only those strains containing the U.S. Reference Factor 6 agglutinogen. Indirect immunofluorescence studies and immunogold electron microscopy demonstrated that these monoclonal antibodies bind to an outer membrane component on serotype-specific strains of B. pertussis. All of the monoclonal antibodies reacted with native or partially assembled type-specific fimbriae but not with monomeric fimbrial subunits as indicated by Western blot (immunoblot) analysis. The fimbrial agglutinogens recognized by the monoclonal antibodies were also uniquely reactive with either U.S. Reference Factor 2 or 6 antiserum (Eldering agglutinogen 2 or 6 polyclonal antiserum) in an indirect ELISA. No cross-reactivity of the monoclonal antibodies with the unrelated fimbriae was observed in any of the comparative immunological studies. Some of the monoclonal antibodies agglutinated certain strains of B. bronchiseptica, suggesting that this closely related species can contain antigenically similar fimbriae. These monoclonal antibodies should prove useful for further structural and functional analysis of Bordetella fimbriae and for studies on the role that these antigens play in prevention of infection and disease. PMID:2903125

  15. Studying red blood cell agglutination by measuring membrane viscosity with optical tweezers

    NASA Astrophysics Data System (ADS)

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

    2007-09-01

    The red blood cell (RBC) viscoelastic membrane contains proteins and glycoproteins embedded in a fluid lipid bilayer that are responsible for cell agglutination. Manipulating RBCs rouleaux with a double optical tweezers, we observed that the cells slide easily one over the others but are strongly connected by their edges. An explanation for this behavior could be the fact that when the cells slide one over the others, proteins are dragged through the membrane. It confers to the movement a viscous characteristic that is dependent of the velocity between the RBCs and justifies why is so easy to slide them apart. Therefore, in a first step of this work, by measuring the force as a function of the relative velocity between two cells, we confirmed this assumption and used this viscous characteristic of the RBC rouleaux to determine the apparent membrane viscosity of the cell. As this behavior is related to the proteins interactions, we can use the apparent membrane viscosity to obtain a better understanding about cell agglutination. Methods related to cell agglutination induced by antigen-antibody interactions are the basis of most of tests used in transfusion centers. Then, in a second step of this work, we measured the apparent membrane viscosity using antibodies. We observed that this methodology is sensitive to different kinds of bindings between RBCs. Better comprehension of the forces and bindings between RBCs could improve the sensibility and specificity of the hemagglutination reactions and also guides the development of new potentiator substances.

  16. Evaluation of latex agglutination and microtube coagulase tests for detection of Staphylococcus aureus.

    PubMed

    Pourshadi, M; Klaas, J

    1984-09-01

    In a blind study, a latex agglutination test (Serostat Staphylococcus, Scott Laboratories) and a microtube coagulase test (Staphase, API) were evaluated for their ability to detect Staphylococcus aureus. Of 289 isolates of catalase-positive, gram-positive cocci, 122 were identified as S. aureus based on positive reactions in at least three of the following tests: tube coagulase, slide coagulase, DNase production, or anaerobic fermentation of mannitol. The latex agglutination test gave positive reactions for all S. aureus isolates and 10 (6%) non-S. aureus isolates. The slide coagulase test was positive for 121 S. aureus isolates and three (2%) non-S. aureus isolates. The microtube coagulase test detected 53, 90, and 98% of the S. aureus isolates after 2, 4, and 24 hr, respectively. In contrast, the conventional tube coagulase test detected 97% of the S. aureus isolates after 2 hr, and 98% after 4 and 24 hr. Two isolates of S. aureus gave negative tube coagulase reactions at 37 degrees C, but positive reactions at room temperature after 24 hr. The combination of tube and slide coagulase tests provided the most reliable results. The slide and tube coagulase tests gave more reliable results than the latex agglutination and microtube coagulase tests, respectively.

  17. 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.

  18. Serological diagnosis of brucellosis in water buffaloes (Bubalus bubalis): comparison among complement fixation, serum agglutination and rose bengal plate test.

    PubMed

    Mathias, L A; Pinto, A A

    1983-12-01

    The results of a comparative study among complement fixation (CFT), plate agglutination (PAT), tube agglutination (TAT) and Rose Bengal plate tests ( RBPT ) to the serodiagnosis of brucellosis in Indian buffaloes are reported. Sera from 212 buffaloes unvaccinated against brucellosis were examined and the CFT was able to reveal significant titres in sera with low agglutinating titres. From 109 sera which did not show agglutination titres in the PAT, four showed complement fixing titre greater than 1 in 200. All the positive sera to the RBPT gave complement fixing titre equal to or greater than 1 in 20. In sera that showed negative result to the RBPT the CFT was able to reveal relatively high titres. From 131 sera negative to the RBPT five showed complement fixing titres greater than 1 in 60.

  19. [Formation of spontaneous and immune rosettes by the cells of the thymus and other formations of the rabbit lymphoid system].

    PubMed

    Grinevich, Iu A

    1975-11-01

    Only T1--RFC (rosette-forming cells) are revealed in the thymus of nonimmunized rabbits. Their number is 2--2.5 times less than in the palatine tonsils, submaxillary lymph nodes and the spleen. T2--RFC are present in these lymphoid formations. There is an increase in the T1--RFC in the thymus after the intravenous immunization of rabbits with sheep erythrocytes. In other lymphoid formations the correlation of the population of cells of the thymus origin altered as a result of increase in the number of T2--RFC. B--RFC accumulated in considerable amounts. Dynamics of T2 and B--RFC accumulation in the lymphoid formations corresponded to the highest antibody titres in the rabbit blood. In the formation of primary immune response the amount of the T1 and T2-RFC in the formations of rabbit lymphoid system depended on the dose of the antigen. PMID:1221703

  20. The modified card agglutination test: an accurate tool for detecting anaplasmosis in Columbian black-tailed deer.

    PubMed

    Howarth, A; Hokama, Y; Amerault, T E

    1976-07-01

    Inoculation of susceptible calves confirmed that the modified card agglutination test accurately detected the anaplasmosis infection status of each of 35 Columbian black-tailed deer (Odocoileus hemionus columbianus). Anaplasma marginale, and specific antibodies, were demonstrated only in calves which received blood from deer that were positive by the card test. The modified card agglutination testing of deer serum was performed in the manner recommended for testing cattle serum with bovine-origin antigen and bovine serum factor.

  1. 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).

  2. 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.

  3. 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

  4. 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.

  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. [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.

  7. 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)

  8. 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.

  9. Brucella melitensis Invades Murine Erythrocytes during Infection

    PubMed Central

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

    2014-01-01

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

  10. 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

  11. 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.

  12. Microscopic agglutination test on captive rattlesnakes : Data on serovars and titers

    PubMed Central

    Rodrigues, T.C.S.; Santos, A.L.Q.; Lima, A.M.C.; Gomes, D.O.; Cardoso, G.F.; Brites, V.L.C.

    2016-01-01

    The microscopic agglutination test (MAT) is considered the “golden standard” leptospirosis serodiagnostic test, but there is little information about it as it pertains to snakes. To fill this information gap, we provide data on serovars and titers of fifty-six Crotalus durissus collilineatus sera samples that tested positive by MAT (10.1016/j.actatropica.2016.02.006 (Rodrigues et al., 2016) [5]). These data are presented in a table, along with a description of the methodology used for sample collection and serologic testing. PMID:27077089

  13. Local hydrocortisone treatment of sperm-agglutinating antibodies in infertile women.

    PubMed

    Ulcová-Gallová, Z; Mráz, L; Plánicková, E; Macků, F; Ulc, I

    1988-01-01

    Local sperm-agglutinating antibodies (LSAA) in the cervical ovulatory mucus may be a cause of primary infertility. A group of 17 infertile women with LSAA treated without effect with artificial insemination and then condom therapy were studied. After hydrocortisone application to the ectocervix for up to four cycles, LSAA disappeared totally in 13 patients; six of them have given birth to babies. No side effects of treatment were observed. Hydrocortisone for local immunosuppression may become a new method of therapy in cervical immunological infertility.

  14. A sperm-agglutinating lectin from seeds of Jack fruit (Artocarpus heterophyllus).

    PubMed

    Namjuntra, P; Muanwongyathi, P; Chulavatnatol, M

    1985-04-30

    A lectin specific for N-acetylgalactosamine was isolated from seed extract of Jack fruit (Artocarpus heterophyllus) by ammonium sulfate precipitation, followed by affinity chromatography on a Affigel-galactosamine-agarose column. The lectin possessed agglutinating activities for human and rat sperm as well as human red blood cells. It was found to have Mr = 62,000 consisting of two dissimilar subunits of Mr = 18,000 and 13,000. It also cross-reacted with an antibody against the lectin of Osage Orange (Maclura pomifera).

  15. 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

  16. Antimicrobial Action and Cell Agglutination by the Eosinophil Cationic Protein Are Modulated by the Cell Wall Lipopolysaccharide Structure

    PubMed Central

    Pulido, David; Moussaoui, Mohammed; Andreu, David; Nogués, M. Victòria

    2012-01-01

    Antimicrobial proteins and peptides (AMPs) are essential effectors of innate immunity, acting as a first line of defense against bacterial infections. Many AMPs exhibit high affinity for cell wall structures such as lipopolysaccharide (LPS), a potent endotoxin able to induce sepsis. Hence, understanding how AMPs can interact with and neutralize LPS endotoxin is of special relevance for human health. Eosinophil cationic protein (ECP) is an eosinophil secreted protein with high activity against both Gram-negative and Gram-positive bacteria. ECP has a remarkable affinity for LPS and a distinctive agglutinating activity. By using a battery of LPS-truncated E. coli mutant strains, we demonstrate that the polysaccharide moiety of LPS is essential for ECP-mediated bacterial agglutination, thereby modulating its antimicrobial action. The mechanism of action of ECP at the bacterial surface is drastically affected by the LPS structure and in particular by its polysaccharide moiety. We have also analyzed an N-terminal fragment that retains the whole protein activity and displays similar cell agglutination behavior. Conversely, a fragment with further minimization of the antimicrobial domain, though retaining the antimicrobial capacity, significantly loses its agglutinating activity, exhibiting a different mechanism of action which is not dependent on the LPS composition. The results highlight the correlation between the protein's antimicrobial activity and its ability to interact with the LPS outer layer and promote bacterial agglutination. PMID:22330910

  17. Antimicrobial action and cell agglutination by the eosinophil cationic protein are modulated by the cell wall lipopolysaccharide structure.

    PubMed

    Pulido, David; Moussaoui, Mohammed; Andreu, David; Nogués, M Victòria; Torrent, Marc; Boix, Ester

    2012-05-01

    Antimicrobial proteins and peptides (AMPs) are essential effectors of innate immunity, acting as a first line of defense against bacterial infections. Many AMPs exhibit high affinity for cell wall structures such as lipopolysaccharide (LPS), a potent endotoxin able to induce sepsis. Hence, understanding how AMPs can interact with and neutralize LPS endotoxin is of special relevance for human health. Eosinophil cationic protein (ECP) is an eosinophil secreted protein with high activity against both Gram-negative and Gram-positive bacteria. ECP has a remarkable affinity for LPS and a distinctive agglutinating activity. By using a battery of LPS-truncated E. coli mutant strains, we demonstrate that the polysaccharide moiety of LPS is essential for ECP-mediated bacterial agglutination, thereby modulating its antimicrobial action. The mechanism of action of ECP at the bacterial surface is drastically affected by the LPS structure and in particular by its polysaccharide moiety. We have also analyzed an N-terminal fragment that retains the whole protein activity and displays similar cell agglutination behavior. Conversely, a fragment with further minimization of the antimicrobial domain, though retaining the antimicrobial capacity, significantly loses its agglutinating activity, exhibiting a different mechanism of action which is not dependent on the LPS composition. The results highlight the correlation between the protein's antimicrobial activity and its ability to interact with the LPS outer layer and promote bacterial agglutination.

  18. Effect of propranolol on normal human erythrocytes.

    PubMed

    Fortier, N L; Snyder, L M; Palek, J; Weiss, E B; Mancini, C; Falcone, J

    1977-01-01

    The present study was undertaken to standardize the effect of propranolol on normal human red cells and thus establish certain parameters enabling us to evaluate propranolol's effect on pathological cells. Normal human erythrocytes lost 40 MEq. of potassium, decreased the intracellular pH by 0.06 units, and shifted the oxyhemoglobin dissociation curve 6.0 mm. Hg to the right in the presence of propranolol. The series of events and magnitude of the response induced by propranolol was time dependent and sensitive to temperature, pH, drug concentration, and erythrocyte concentration. Calcium was an absolute requirement for maximal propranolol action with simultaneous incorporation of trace amounts of radioactive calcium into the cell. Chelation of calcium with EDTA or EGTA inhibited the response to propranolol.

  19. Determinants of Erythrocyte Hydration In Current Opinion in Hematology

    PubMed Central

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

    2012-01-01

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

  20. Phase separation in frozen erythrocyte membrane preparations.

    PubMed

    Finean, J B; Hutchinson, A; Mills, D

    1985-10-01

    The reversible formation of a lipid-like phase in frozen preparations of erythrocyte membranes has been studied by X-ray diffraction and by electron microscopy of freeze-fracture replicas. The observations provide strong evidence for lateral migration or displacement at specific temperatures of intra-membrane particles. This creates large areas of particle-free membranes which fracture preferentially so as to dominate the freeze-fracture image.

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

    PubMed

    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

  2. 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.

  3. 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

  4. Homogeneous agglutination assay based on micro-chip sheathless flow cytometry.

    PubMed

    Ma, Zengshuai; Zhang, Pan; Cheng, Yinuo; Xie, Shuai; Zhang, Shuai; Ye, Xiongying

    2015-11-01

    Homogeneous assays possess important advantages that no washing or physical separation is required, contributing to robust protocols and easy implementation which ensures potential point-of-care applications. Optimizing the detection strategy to reduce the number of reagents used and simplify the detection device is desirable. A method of homogeneous bead-agglutination assay based on micro-chip sheathless flow cytometry has been developed. The detection processes include mixing the capture-probe conjugated beads with an analyte containing sample, followed by flowing the reaction mixtures through the micro-chip sheathless flow cytometric device. The analyte concentrations were detected by counting the proportion of monomers in the reaction mixtures. Streptavidin-coated magnetic beads and biotinylated bovine serum albumin (bBSA) were used as a model system to verify the method, and detection limits of 0.15 pM and 1.5 pM for bBSA were achieved, using commercial Calibur and the developed micro-chip sheathless flow cytometric device, respectively. The setup of the micro-chip sheathless flow cytometric device is significantly simple; meanwhile, the system maintains relatively high sensitivity, which mainly benefits from the application of forward scattering to distinguish aggregates from monomers. The micro-chip sheathless flow cytometric device for bead agglutination detection provides us with a promising method for versatile immunoassays on microfluidic platforms.

  5. 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.

  6. Agglutinated tests in post-Sturtian cap carbonates of Namibia and Mongolia

    NASA Astrophysics Data System (ADS)

    Bosak, T.; Lahr, D. J. G.; Pruss, S. B.; Macdonald, F. A.; Dalton, L.; Matys, E.

    2011-08-01

    Paleomagnetic data suggest that the early Cryogenian (Sturtian) glaciation extended to sea level at low latitude. The impact of this dramatic environmental change on biota, and the composition of ecosystems in the immediate aftermath of the Sturtian glaciation remain virtually unknown. Here we report the discovery of abundant agglutinated tests in organic-rich carbonates directly overlying Sturtian glacial deposits from two different paleocontinents: the Rasthof Formation of the Congo craton in northern Namibia and the Tsagaan Oloom Formation of the Dzabkhan terrane in Mongolia. The most abundant tests preserve morphological and compositional characters consistent with those found in at least two different families of modern lobose testate amoebae (Amoebozoa), a group of heterotrophic microbial eukaryotes. The presence of spatially and compositionally variable clay minerals, quartz and microcline on the test walls is a signature of widespread biological agglutination. The post-glacial fossil assemblages differ from the most common pre-Sturtian vase-shaped fossil testate amoebae, perhaps as a result of different preservational mechanisms or of the appearance of new forms after the glaciation. The apparent local abundance of eukaryotic body fossils in the post-Sturtian carbonates suggests that the Cryogenian limestones and dolostones may host a currently unexplored fossil record of modern eukaryotes.

  7. Seroprevalence of Mycoplasma pneumoniae in HIV-infected patients using a microparticle agglutination test.

    PubMed

    Shankar, Esaki Muthu; Kumarasamy, Nagalingeswaran; Balakrishnan, Pachamuthu; Vengatesan, A; Kownhar, Hayath; Solomon, Suniti; Rao, Usha Anand

    2006-06-01

    Mycoplasma pneumoniae is increasingly recognized as a common and important pathogen in community settings, and is responsible for various pulmonary and extrapulmonary conditions in the normal population. However, the seroepidemiology of acute M. pneumoniae infection in HIV-infected individuals is still unclear worldwide. This study examined the seroprevalence of antibodies to M. pneumoniae in HIV-infected patients admitted with respiratory complaints at a tertiary AIDS care centre in Chennai, India. A commercial gelatin microparticle agglutination test (Serodia-Myco II, Fujirebio) was used for the determination of antibodies against M. pneumoniae in acute serum specimens. Of the 200 HIV-infected patients with underlying pulmonary conditions tested, 34 (17 % positivity; 95 % CI 12-23 %) had antibodies specific to M. pneumoniae, while among the 40 patients with no underlying pulmonary symptoms, five (12.5 % positivity; 95 % CI 4-27 %) had evidence of anti-M. pneumoniae antibody. This shows that the incidence of M. pneumoniae seropositivity is greater in patients with underlying pulmonary complaints. Most positive titres were found in the age group 28-37 years in the symptomatic and symptom-free groups (64.7 and 60 %, respectively). The positive titres ranged from 40 to >20 480. High titres (> or =320) were found in 10 out of the 39 patients (25.6 %). This seroprevalence study reports a 16.2 % prevalence of M. pneumoniae infections in HIV-infected patients by a particle agglutination test.

  8. Erythrocyte shape simulation by numerical optimization.

    PubMed

    Grebe, R; Zuckermann, M J

    1990-01-01

    In a recent paper we examined the morphology of erythrocytes in terms of the mean mean curvature (MMC) of their cell membranes. A computer simulation of these shapes based on the different geometries showed that the MMC increased from the sphero-stomatocyte to the spheroechinocyte via the discocyte. In this work we extend this analysis by using a numerical optimization method based on importance sampling and the principle of adiabatic cooling. The erythrocyte membrane is treated as a single closed fluid lamina exhibiting viscoelastic characteristics. The energy function of the lamina includes the following terms: (i) Curvature-elastic energy terms which depend on both local and global curvature. (ii) A term describing the compression elasticity of the lamina. (iii) A term which depends on the volume of the cell and which is related to the osmotic pressure across the membrane. In the simulation the cell is assumed to have axial symmetry and it can therefore be described by a finite set of conic sections. So far we have been able to obtain an energy minimum corresponding to a discocyte shape using a sphere as the initial configuration. Our results therefore imply that the well-known sequence of erythrocyte shapes could solely be governed by the above mentioned properties of an ideal fluid forming a closed singly connected lamina.

  9. Discovery of Novel Plasmodium falciparum Pre-Erythrocytic Antigens for Vaccine Development

    PubMed Central

    Aguiar, Joao C.; Bolton, Jessica; Wanga, Joyce; Sacci, John B.; Iriko, Hideyuki; Mazeika, Julie K.; Han, Eun-Taek; Limbach, Keith; Patterson, Noelle B.; Sedegah, Martha; Cruz, Ann-Marie; Tsuboi, Takafumi; Hoffman, Stephen L.; Carucci, Daniel; Hollingdale, Michael R.; Villasante, Eileen D.; Richie, Thomas L.

    2015-01-01

    Background Nearly 100% protection against malaria infection can be achieved in humans by immunization with P. falciparum radiation-attenuated sporozoites (RAS). Although it is thought that protection is mediated by T cell and antibody responses, only a few of the many pre-erythrocytic (sporozoite and liver stage) antigens that are targeted by these responses have been identified. Methodology Twenty seven P. falciparum pre-erythrocytic antigens were selected using bioinformatics analysis and expression databases and were expressed in a wheat germ cell-free protein expression system. Recombinant proteins were recognized by plasma from RAS-immunized subjects, and 21 induced detectable antibody responses in mice and rabbit and sera from these immunized animals were used to characterize these antigens. All 21 proteins localized to the sporozoite: five localized to the surface, seven localized to the micronemes, cytoplasm, endoplasmic reticulum or nucleus, two localized to the surface and cytoplasm, and seven remain undetermined. PBMC from RAS-immunized volunteers elicited positive ex vivo or cultured ELISpot responses against peptides from 20 of the 21 antigens. Conclusions These T cell and antibody responses support our approach of using reagents from RAS-immunized subjects to screen potential vaccine antigens, and have led to the identification of a panel of novel P. falciparum antigens. These results provide evidence to further evaluate these antigens as vaccine candidates. Trial Registration ClinicalTrials.gov NCT00870987 ClinicalTrials.gov NCT00392015 PMID:26292257

  10. Proposed method for agglutinating antibody titer analysis and its use as indicator of acquired immunity in pacu, Piaractus mesopotamicus.

    PubMed

    Biller-Takahashi, J D; Montassier, H J; Takahashi, L S; Urbinati, E C

    2014-02-01

    Antibody can be assessed by agglutinating antibody titer which is a quantitative measure of circulating antibodies in serum from fish previously immunized. The antibody evaluation has been performed with different fish species, and is considered a reliable method that can be applied to confirm several hypothesis regarding acquired immunity, even in conjunction with precise methods to describe immune mechanisms. In order to provide appropriate analytical methods for future studies on the specific immune system of native fish, the present study standardized on assay to measure the serum agglutinating antibody titer produced after immunization with inactivated A. hydrophila and levamisole administration in pacu. It was possible to determine the agglutinating antibodies titer in a satisfactorily way in pacu immunized with inactive A. hydrophila, and the highest titers were observed on fish fed with levamisole.

  11. Spectral Markers of Erythrocytes on Solid Substrate

    NASA Astrophysics Data System (ADS)

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

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

  12. 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.

  13. 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. PMID:8085400

  14. Uric acid protects erythrocytes from ozone-induced changes

    SciTech Connect

    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.

  15. Effect of curcumin on LDL oxidation in vitro, and lipid peroxidation and antioxidant enzymes in cholesterol fed rabbits.

    PubMed

    Mahfouz, Mohamedain M; Zhou, Qi; Kummerow, Fred A

    2011-11-01

    In this study we examined the antioxidant effect of curcumin on lipid oxidation in vitro and in vivo. In vitro, curcumin at 5 microgM concentration completely prevented low-density lipoprotein (LDL) oxidation by CuS0(4), indicating that curcumin is an effective antioxidant in vitro. In vivo, feeding a pure cholesterol (PC)-rich diet to rabbits significantly increased the plasma and liver lipids as well as thiobarbituric acid reactive substances (TBARS) levels. Addition of curcumin to the PC diet did not show any effect on either plasma lipid and TBARS or liver lipids. Liver TBARS tended to decrease but that decrease was not significant. Erythrocyte glutathione peroxidase (GSH-Px) activity was significantly decreased while catalase activity was significantly increased in rabbits fed a PC diet. The addition of curcumin to a PC diet did not show any significant effect on erythrocyte enzyme activities compared to the rabbits fed a PC diet. The liver GSH-Px and catalase activities were significantly decreased in rabbits fed a PC diet, but the addition of curcumin to the PC diet enhanced the liver GSH-Px activity, which became nonsignificantly different from the control group. These results were discussed considering that curcumin may not be well absorbed and it did not reach a level high enough in vivo to overcome the severe hypercholesterolemia and oxidative stress produced by the PC-rich diet.

  16. Hepatic or splenic targeting of carrier erythrocytes: a murine model

    SciTech Connect

    Zocchi, E.; Guida, L.; Benatti, U.; Canepa, M.; Borgiani, L.; Zanin, T.; De Flora, A.

    1987-10-01

    Carrier mouse erythrocytes, i.e., red cells, subjected to a dialysis technique involving transient hypotonic hemolysis and isotonic resealing were treated in vitro in three different ways: (a) energy depletion by exposure for 90 min at 42 degrees C; (b) desialylation by incubation with neuroaminidase; and (c) oxidative stress by incubation with H/sub 2/O/sub 2/ and NaN3. Procedure (c) afforded maximal damage, as shown by analysis of biochemical properties of the treated erythrocytes. Reinfusion in mice of the variously manipulated erythrocytes following their /sup 51/Cr labeling showed extensive fragilization as indicated by rapid clearance of radioactivity from the circulation. Moreover, both the energy-depleted and the neuraminidase-treated erythrocytes showed a preferential liver uptake, reaching 50 and 75%, respectively, within 2 h. On the other hand, exposure of erythrocytes to the oxidant stress triggered a largely splenic removal, accounting for almost 40% of the reinjected cells within 4 h. Transmission electron microscopy of liver from mice receiving energy-depleted erythrocytes demonstrated remarkable erythrocyte congestion within the sinusoids, followed by hyperactivity of Kupffer cells and by subsequent thickening of the perisinusoidal Disse space. Concomitantly, levels of serum transaminase activities were moderately increased. Each of the three procedures of manipulation of carrier erythrocytes may prove applicable under conditions where selective targeting of erythrocyte-encapsulated chemicals and drugs to either the liver or the spleen has to be achieved.

  17. 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.

  18. Erythrophagocytosis by Liver Macrophages (Kupffer Cells) Promotes Oxidative Stress, Inflammation, and Fibrosis in a Rabbit Model of Steatohepatitis

    PubMed Central

    Otogawa, Kohji; Kinoshita, Kohji; Fujii, Hideki; Sakabe, Masahide; Shiga, Ryoko; Nakatani, Kazuki; Ikeda, Kazuo; Nakajima, Yuji; Ikura, Yoshihiro; Ueda, Makiko; Arakawa, Tetsuo; Hato, Fumihiko; Kawada, Norifumi

    2007-01-01

    Nonalcoholic steatohepatitis (NASH) is a progressive fibrotic disease, the pathogenesis of which has not been fully elucidated. Here, we report a molecular aspect of this disease elucidated using rabbits fed a cholesterol-rich high-fat diet and exhibiting insulin resistance. The liver in this model showed steatohepatitis with fibrosis and high mRNA expression for some cytokines, heme oxygenase-1, transforming growth factor-β1, and collagen α1(I). Erythrocytes isolated from the model showed marked fragility and the externalization of phosphatidylserine (PS) on the outer leaflet of the membrane and were frequently engulfed by Kupffer cells/macrophages in the hepatic sinusoids. Expression of milk fat globule-epidermal growth factor (EGF)-factor 8, a PS-binding protein, was augmented in the liver. In culture, RAW 264.7 cells engulfed erythrocytes oxidized by tert-butyl hydroperoxide, a process that was inhibited by anti-milk fat globule-EGF-factor 8 antibody. In addition, PS-positive erythrocytes appeared entrapped in the model liver in ex vivo perfusion experiments. Finally, in specimens from NASH patients, the aggregation of erythrocytes in inflammatory hepatic sinusoids was notable. These results indicate that the engulfment of PS-externalized, apoptotic signal-positive, erythrocytes by hepatic macrophages may lead to the deposition of iron derived from hemoglobin in the liver and be involved in the pathogenesis of steatohepatitis. PMID:17322381

  19. 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

  20. Seroprevalence of bovine leptospiral antibodies by microscopic agglutination test in Southeast of Iran

    PubMed Central

    Khalili, Mohammad; Sakhaee, Ehsanollah; Aflatoonian, Mohammad Reza; Abdollahpour, Gholamreza; Tabrizi, Saeed Sattari; Damaneh, Elham Mohammadi; Hossini-nasab, Sajad

    2014-01-01

    Objective To evaluate serological findings of bovine leptospirosis which is a zoonotic disease with worldwide distribution caused by Leptospira interrogans. Methods One hundred and sixty seven sera were collected from 9 commercial dairy herds in jiroft suburbs, from July to October 2011. Microscopic agglutination test (MAT) was used to evaluates serological findings of bovine leptospirosis in Jiroft suburb dairy farms, Kerman province, Iran. Results Antibodies were found by MAT at least against one serovar of Leptospira interrogans in 29 samples (17.36%) among 167 sera at a dilution 1:100 or higher, and Leptospira pomona was the most prevalent serovar. Positive titers against more than one serovar were detected in 6 sera of the positive samples. Conclusion This study is the first report of leptospirosis in Southeast Iran and showed that Leptospira pomona was the most and Leptospira icterohaemorrhagiae the least prevalent serovars in Southeast Iran. PMID:25182718

  1. Pneumococcal antigen detection in cerebrospinal fluid: a comparative study on counter immunoelectrophoresis, latex agglutination and coagglutination.

    PubMed

    Rai, G P; Zachariah, K; Sharma, R; Phadke, S; Belapurkar, K M

    2003-07-01

    The sensitivity, specificity, accuracy and predictive values of counter immunoelectrophoresis (CIE), latex agglutination (LA) and coagglutination (CoAg) tests were compared for detection of pneumococcal antigen in cerebrospinal fluid (CSF) of patients suspected of meningitis. A total of 95 CSF samples comprising 15 culture proven, 47 clinically suspected but culture negative cases of meningitis and 33 controls were screened by above tests. Among three tests, LA was found to have high sensitivity and moderately high negative predictive value than CIE and CoAg tests. However, CIE had slightly better specificity than LA and CoAg tests. Accuracywise CIE and LA tests were comparable than CoAg test. CIE and LA tests had high positive predictive value than CoAg test.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. Blood viscosity: influence of erythrocyte aggregation.

    PubMed

    Chien, S; Usami, S; Dellenback, R J; Gregersen, M I; Nanninga, L B; Guest, M M

    1967-08-18

    The addition of purified canine or bovine fibrinogen to suspensions of canine erythocytes in Ringer solution caused an increase in viscosity and the formation of aggregates of erythocytes. Both of these effects became increasingly pronounced as the fibrinogen concentration was raised, and they approached plateaus with 1 gram of fibrinogen per 100 milliliters. An increase in shear rate (or shear stress) reduced both the effect on viscosity and the aggregate size. The data suggest that fibrinogen causes an increase in blood viscosity and a departure from Newtonian behavior by interacting with erythrocytes to form cell aggregates which can be dispersed by shear stress. PMID:17842794

  7. Cflec-5, a pattern recognition receptor in scallop Chlamys farreri agglutinating yeast Pichia pastoris.

    PubMed

    Zhang, Huan; Kong, Pengfei; Wang, Lingling; Zhou, Zhi; Yang, Jialong; Zhang, Ying; Qiu, Limei; Song, Linsheng

    2010-07-01

    C-type lectins are a superfamily of carbohydrate-recognition proteins which play crucial roles as pattern recognition receptors (PRRs) in the innate immunity. In this study, the full-length cDNA of a C-type lectin was cloned from scallop Chlamys farreri (designated as Cflec-5) by expression sequence tag (EST) analysis and rapid amplification of cDNA ends (RACE) approach. The full-length cDNA of Cflec-5 was of 1412 bp. The open reading frame encoded a polypeptide of 153 amino acids, including a signal sequence and a conserved carbohydrate-recognition domain with the EPN motif determining the mannose-binding specificity. The deduced amino acid sequence of Cflec-5 showed high similarity to members of C-type lectin superfamily. The quantitative real-time PCR was performed to investigate the tissue distribution of Cflec-5 mRNA and its temporal expression profiles in hemocytes post pathogen-associated molecular patterns (PAMPs) stimulation. In healthy scallops, the Cflec-5 mRNA was mainly detected in gill and mantle, and marginally in other tissues. The mRNA expression of Cflec-5 could be significantly induced by lipopolysaccharide (LPS) and glucan stimulation and reached the maximum level at 6 h and 12 h, respectively. But its expression level did not change significantly during peptidoglycan (PGN) stimulation. The function of Cflec-5 was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli Rosetta Gami (DE3). The recombinant Cflec-5 agglutinated Pichia pastoris in a calcium-independent way. The agglutinating activity could be inhibited by d-mannose, LPS and glucan, but not by d-galactose or PGN. These results collectively suggested that Cflec-5 was involved in the innate immune response of scallops and might contribute to nonself-recognition through its interaction with various PAMPs. PMID:20211738

  8. [Evaluation of gelatin particle agglutination method for detection of Treponema pallidum antibody].

    PubMed

    Deguchi, M; Hosotsubo, H; Yamashita, N; Ohmine, T; Asari, S

    1994-10-01

    Treponema pallidum hemagglutination (HA) is one of the most frequently used methods for the detection of Treponema pallidum (T. pallidum) antibodies. Recently, an innovative agglutination method using artificial carriers was newly developed, and is now available as a routine method. In order to compare the newly developed particle agglutination (PA) method (FUJIREBIO INC.) with the conventional HA method, T. pallidum antibody titers of numerous sera were measured by respective methods. In the stability study, reconstituted reagent was stable for at least three weeks. Sample inactivation (56 degrees C/30 min) demonstrated no effect on the test results. Among 800 sera, 132 (16.6%) positives (+), 633 (79.1%) negatives (-) and (4.3%) indeterminates (+) were obtained by HA method. Meanwhile, 144 (18.0%) positives (+), 627 (78.4%) negatives (-) and 29 (3.6%) indeterminates (+) were obtained by PA method. The correlation between PA and HA method was 97.8%, and the antibody titers obtained by PA method showed good correlation with HA method. Those samples which showed discrepancy between PA and HA method in the above study were further examined with fluorescent treponemal antibody-absorption (FTA-ABS) method. The results obtained from FTA-ABS method were almost consistent with those obtained from PA method. For respective syphilis patients in stage I and II, antibody titer was monitored by HA, PA and RPR method. The results indicated that changes in antibody titer obtained from PA method was approximately the same as the titer changes obtained from RPR method. Namely, PA method detected the presence of IgM earlier than HA method.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. 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.

  10. 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.

  11. Development of a latex agglutination test with recombinant variant surface glycoprotein for serodiagnosis of surra.

    PubMed

    Rogé, S; Baelmans, R; Claes, F; Lejon, V; Guisez, Y; Jacquet, D; Büscher, P

    2014-10-15

    Serodiagnosis of surra is commonly performed with the CATT/Trypanosoma evansi direct agglutination test. This antibody detection test is based on lyophilised bloodstream form trypanosomes propagated in rats and presenting the predominant variant surface glycoprotein (VSG) RoTat 1.2 on their surface. Recently, the N-terminal fragment of VSG RoTat 1.2 has been expressed as a recombinant protein in the yeast Pichia pastoris and showed diagnostic potential in ELISA. This recombinant antigen has now been incorporated in a latex agglutination test, the rLATEX/T. evansi. In this study, we compared the diagnostic accuracy of rLATEX/T. evansi and CATT/T. evansi with immune trypanolysis (TL) as reference test on a total of 1717 sera from camels, horses, bovines, water buffaloes, dogs and sheep. The rLATEX/T. evansi displayed a slightly better agreement with TL than CATT/T. evansi (kappa [κ] respectively 0.84 and 0.72). The sensitivities of rLATEX/T. evansi (84.2%, 95% CI 80.8-87.1) and CATT/T. evansi (84.0%, 95% CI 80.6-87.0) were similar, but rLATEX/T. evansi was significantly more specific (97.7%, 95% CI 96.7-98.4) than CATT/T. evansi (89.4%; 95% CI 87.6-91.1). We consider the rLATEX/T. evansi an alternative for the CATT/T. evansi, with the advantage that the use of a purified recombinant antigen leads to a more standardised diagnostic test with an improved specificity. Moreover, it eliminates the use of laboratory animals and can be easily scaled-up, e.g. in biofermentors.

  12. Study of agglutination of mouse mammary carcinoma (FM3A) cell induced by egg agglutinin of Rana catesbiana. II. Phytohemagglutinin P and protamine.

    PubMed

    Takeda, S; Kubota, K; Endo, Y; Matsuzawa, T

    1981-01-01

    When mouse mammary carcinoma (FM3A) cells were treated with egg agglutinin of Rana catesbiana for 15 min at 25 degrees C, percent total particle number of both cell aggregates and single cells was in direct proportion to the cell electrophoretic mobility. Phytohemagglutinin P mediated agglutination proceeded with biphasic kinetics: the higher the concentration of phytohemagglutinin P the shorter was the lag period between the first and second stages of agglutination. In protamine-mediated agglutination, the percent total particle number was reduced at low concentrations, while the electrophoretic mobility reduced only at high concentrations. Agglutinating and cytotoxic activities of these three reagents were in an intimate relation: the higher the agglutinating activity, the greater was their cytotoxic activity. PMID:6969671

  13. Agglutination of Histoplasma capsulatum by IgG monoclonal antibodies against Hsp60 impacts macrophage effector functions.

    PubMed

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

    2011-02-01

    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.

  14. 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

  15. 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...

  16. Modeling of Virion Collisions in Cervicovaginal Mucus Reveals Limits on Agglutination as the Protective Mechanism of Secretory Immunoglobulin A

    PubMed Central

    Chen, Alex; McKinley, Scott A.; Shi, Feng; Wang, Simi; Mucha, Peter J.; Harit, Dimple; Forest, M. Gregory; Lai, Samuel K.

    2015-01-01

    Secretory immunoglobulin A (sIgA), a dimeric antibody found in high quantities in the gastrointestinal mucosa, is broadly associated with mucosal immune protection. A distinguishing feature of sIgA is its ability to crosslink pathogens, thereby creating pathogen/sIgA aggregates that are too large to traverse the dense matrix of mucin fibers in mucus layers overlying epithelial cells and consequently reducing infectivity. Here, we use modeling to investigate this mechanism of “immune exclusion” based on sIgA-mediated agglutination, in particular the potential use of sIgA to agglutinate HIV in cervicovaginal mucus (CVM) and prevent HIV transmission. Utilizing reported data on HIV diffusion in CVM and semen, we simulate HIV collision kinetics in physiologically-thick mucus layers–a necessary first step for sIgA-induced aggregation. We find that even at the median HIV load in semen of acutely infected individuals possessing high viral titers, over 99% of HIV virions will penetrate CVM and reach the vaginal epithelium without colliding with another virion. These findings imply that agglutination is unlikely to be the dominant mechanism of sIgA-mediated protection against HIV or other sexually transmitted pathogens. Rather, we surmise that agglutination is most effective against pathogens either present at exceedingly high concentrations or that possess motility mechanisms other than Brownian diffusion that significantly enhance encounter rates. PMID:26132216

  17. THE RABBIT ZYGOTE

    PubMed Central

    Gulyas, Bela J.

    1972-01-01

    The formation of the blastomere nucleus was examined in the rabbit zygote with the electron microscope. In late anaphase the chromosomes are bare and vesicles of the smooth endoplasmic reticulum are numerous in the vicinity of the chromosomes. In early telophase individual chromosomes attain their own nuclear envelope and they are called karyomeres. The envelope of the karyomeres contains small gaps within it at several places where the chromatin is exposed to the cytoplasm. Nuclear pores are also observed. In the cytoplasm short annulate lamellae appear adjacent to the karyomeres, and clusters of punctate substance are also present. From early telophase onward the karyomeres extend pseudopod-like structures, called karyopods, which extend toward other karyomeres or karyopods, and consequently fuse together and serve as chromosomal bridges. Eventually all of the karyomeres fuse into a dense nucleus and decondensation of the chromosomes occurs. PMID:4676367

  18. Metabolic properties of low ATP erythrocytes of the monotremes.

    PubMed

    Kim, H D; Zeidler, R B; Sallis, J; Nicol, S; Isaacks, R E

    1984-02-13

    The erythrocytes of the monotremes, having a trace amount of ATP, can metabolize glucose to lactate at a rate comparable to human and other mammalian erythrocytes. The echidna energy metabolism is unique in that adenosine can stimulate glycolytic carbon flow, resulting in a nearly 20-fold net synthesis of ATP.

  19. 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.

  20. Histamine increases sickle erythrocyte adherence to endothelium.

    PubMed

    Wagner, Matthew C; Eckman, James R; Wick, Timothy M

    2006-02-01

    Complications of sickle cell anaemia include vascular occlusion triggered by the adherence of sickle erythrocytes to endothelium in the postcapillary venules. Adherence can be promoted by inflammatory mediators that induce endothelial cell adhesion molecule expression and arrest flowing erythrocytes. The present study characterised the effect of histamine stimulation on the kinetics of sickle cell adherence to large vessel and microvascular endothelium under physiological flow. Increased sickle cell adherence was observed within minutes of endothelial activation by histamine and reached a maximum value within 30 min. At steady state, sickle cell adherence to histamine-stimulated endothelium was 47 +/- 4 adherent cells/mm(2), 2.6-fold higher than sickle cell adherence to unstimulated endothelial cells. Histamine-induced sickle cell adherence occurred rapidly and transiently. Studies using histamine receptor agonists and antagonists suggest that histamine-induced sickle cell adhesion depends on simultaneous stimulation of the H(2) and H(4) histamine receptors and endothelial P-selectin expression. These data show that histamine release may promote sickle cell adherence and vaso-occlusion. In vivo histamine release should be studied to determine its role in sickle complications and whether blocking of specific histamine receptors may prevent clinical complications or adverse effects from histamine release stimulated by opiate analgesic treatment.

  1. Binding characteristics of swine erythrocyte insulin receptors

    SciTech Connect

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

    1985-09-01

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

  2. 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. PMID:7599974

  3. 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.

  4. Plasmodium falciparum Secretome in Erythrocyte and Beyond

    PubMed Central

    Soni, Rani; Sharma, Drista; Bhatt, Tarun K.

    2016-01-01

    Plasmodium falciparum is the causative agent of deadly malaria disease. It is an intracellular eukaryote and completes its multi-stage life cycle spanning the two hosts viz, mosquito and human. In order to habituate within host environment, parasite conform several strategies to evade host immune responses such as surface antigen polymorphism or modulation of host immune system and it is mediated by secretion of proteins from parasite to the host erythrocyte and beyond, collectively known as, malaria secretome. In this review, we will discuss about the deployment of parasitic secretory protein in mechanism implicated for immune evasion, protein trafficking, providing virulence, changing permeability and cyto-adherence of infected erythrocyte. We will be covering the possibilities of developing malaria secretome as a drug/vaccine target. This gathered information will be worthwhile in depicting a well-organized picture for host-pathogen interplay during the malaria infection and may also provide some clues for the development of novel anti-malarial therapies. PMID:26925057

  5. 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

  6. A low cost and high throughput magnetic bead-based immuno-agglutination assay in confined droplets.

    PubMed

    Teste, Bruno; Ali-Cherif, Anaïs; Viovy, Jean Louis; Malaquin, Laurent

    2013-06-21

    Although passive immuno-agglutination assays consist of one step and simple procedures, they are usually not adapted for high throughput analyses and they require expensive and bulky equipment for quantitation steps. Here we demonstrate a low cost, multimodal and high throughput immuno-agglutination assay that relies on a combination of magnetic beads (MBs), droplets microfluidics and magnetic tweezers. Antibody coated MBs were used as a capture support in the homogeneous phase. Following the immune interaction, water in oil droplets containing MBs and analytes were generated and transported in Teflon tubing. When passing in between magnetic tweezers, the MBs contained in the droplets were magnetically confined in order to enhance the agglutination rate and kinetics. When releasing the magnetic field, the internal recirculation flows in the droplet induce shear forces that favor MBs redispersion. In the presence of the analyte, the system preserves specific interactions and MBs stay in the aggregated state while in the case of a non-specific analyte, redispersion of particles occurs. The analyte quantitation procedure relies on the MBs redispersion rate within the droplet. The influence of different parameters such as magnetic field intensity, flow rate and MBs concentration on the agglutination performances have been investigated and optimized. Although the immuno-agglutination assay described in this work may not compete with enzyme linked immunosorbent assay (ELISA) in terms of sensitivity, it offers major advantages regarding the reagents consumption (analysis is performed in sub microliter droplet) and the platform cost that yields to very cheap analyses. Moreover the fully automated analysis procedure provides reproducible analyses with throughput well above those of existing technologies. We demonstrated the detection of biotinylated phosphatase alkaline in 100 nL sample volumes with an analysis rate of 300 assays per hour and a limit of detection of 100 pM.

  7. Evaluation of six agglutination tests for Staphylococcus aureus identification depending upon local prevalence of meticillin-resistant S. aureus (MRSA).

    PubMed

    Weist, Klaus; Cimbal, Ann-Katrin; Lecke, Christoph; Kampf, Günter; Rüden, Henning; Vonberg, Ralf-Peter

    2006-03-01

    Most routine laboratory detection of Staphylococcus aureus isolates is based on rapid agglutination test systems. Failure of agglutination assays to identify meticillin-resistant S. aureus strains (MRSA) has been demonstrated. The aim of this study was to evaluate six commercially available agglutination tests for the detection of meticillin-sensitive S. aureus (MSSA) and mecA-positive MRSA strains. The Dry Spot Staphytect Plus test (Oxoid), the Pastorex Staph Plus test (Bio-Rad), the Slidex Staph-Kit and Slidex Staph Plus test (bioMérieux), the Staphaurex Plus test (Remel) and the Staphylase Test (Oxoid) were used. As determined by pulsed field gel electrophoresis, 52 distinct MRSA strains from five countries, 83 MSSA strains and 150 coagulase-negative staphylococci were included. Species identification and determination of susceptibility patterns were performed using colony morphology, Gram stain, catalase testing, tube coagulase testing, DNase testing, mannitol fermentation, susceptibility testing towards oxacillin by Etest, coagulase gene PCR, fibrinogen receptor gene PCR and PCR of the mecA gene. Sensitivity of the agglutination tests ranged from 82.7 to 100.0 % for MRSA strains and 92.8 to 100.0 % for MSSA strains, respectively. Specificity of the test systems ranged from 91.3 to 99.1 %. None of the six agglutination assays produced correct reactions for all staphylococci tested. Only the Dry Spot Staphytect Plus test correctly identified all 52 MRSA strains. For the other tests kits, sensitivity of MRSA detection was lower than for MSSA isolates. Depending upon the local MRSA prevalence and the parameter of interest (sensitivity or specificity), these test systems may be useful for routine diagnostic purposes.

  8. 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.

  9. 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.

  10. Encapsulation of thiosulfate: cyanide sulfurtransferase by mouse erythrocytes

    SciTech Connect

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

    1986-03-30

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

  11. 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.

  12. Effect of solution non-ideality on erythrocyte volume regulation.

    PubMed

    Levin, R L; Cravalho, E G; Huggins, C E

    1977-03-01

    A non-ideal, hydrated, non-dilute pseudo-binary salt-protein-water solution model of the erythrocyte intracellular solution is presented to describe the osmotic behavior of human erythrocytes. Existing experimental activity data for salts and proteins in aqueous solutions are used to formulate van Laar type expressions for the solvent and solute activity coefficients. Reasonable estimates can therefore be made of the non-ideality of the erythrocyte intracellular solution over a wide range of osmolalities. Solution non-ideality is shown to affect significantly the degree of solute polarization within the erythrocyte intracellular solution during freezing. However, the non-ideality has very little effect upon the amount of water retained within erythrocytes cooled at sub-zero temperatures. PMID:16250333

  13. Microsphiltration: a microsphere matrix to explore erythrocyte deformability.

    PubMed

    Lavazec, Catherine; Deplaine, Guillaume; Safeukui, Innocent; Perrot, Sylvie; Milon, Geneviève; Mercereau-Puijalon, Odile; David, Peter H; Buffet, Pierre

    2013-01-01

    The altered deformability of erythrocytes infected with Plasmodium falciparum is central in malaria -pathogenesis, as it influences the hemodynamic properties of the infected cell and its retention in the spleen. Exported parasite proteins, as well as the shape and volume of the parasite itself, influence the deformability of the infected erythrocyte. To explore changes in erythrocyte deformability, we have developed a new method, called microsphiltration, based on filtration of erythrocytes through a mixture of metal microspheres that mimic the geometry of inter-endothelial splenic slits. As P. falciparum develops in its host cell, the retention rates observed in microspheres correlate with the progressive decrease of erythrocyte deformability and with the retention rates in the spleen. The yields of microsphiltration separation allow for molecular analyses of subpopulations with distinct mechanical phenotypes.

  14. Small, clonally variant antigens expressed on the surface of the Plasmodium falciparum-infected erythrocyte are encoded by the rif gene family and are the target of human immune responses.

    PubMed

    Fernandez, V; Hommel, M; Chen, Q; Hagblom, P; Wahlgren, M

    1999-11-15

    Disease severity in Plasmodium falciparum infections is a direct consequence of the parasite's efficient evasion of the defense mechanisms of the human host. To date, one parasite-derived molecule, the antigenically variant adhesin P. falciparum erythrocyte membrane protein 1 (PfEMP1), is known to be transported to the infected erythrocyte (pRBC) surface, where it mediates binding to different host receptors. Here we report that multiple additional proteins are expressed by the parasite at the pRBC surface, including a large cluster of clonally variant antigens of 30-45 kD. We have found these antigens to be identical to the rifins, predicted polypeptides encoded by the rif multigene family. These parasite products, formerly called rosettins after their identification in rosetting parasites, are prominently expressed by fresh isolates of P. falciparum. Rifins are immunogenic in natural infections and strain-specifically recognized by human immune sera in immunoprecipitation of surface-labeled pRBC extracts. Furthermore, human immune sera agglutinate pRBCs digested with trypsin at conditions such that radioiodinated PfEMP1 polypeptides are not detected but rifins are detected, suggesting the presence of epitopes in rifins targeted by agglutinating antibodies. When analyzed by two-dimensional electrophoresis, the rifins resolved into several isoforms in the pI range of 5.5-6.5, indicating molecular microheterogeneity, an additional potential novel source of antigenic diversity in P. falciparum. Prominent polypeptides of 20, 22, 76-80, 140, and 170 kD were also detected on the surfaces of pRBCs bearing in vitro-propagated or field-isolated parasites. In this report, we describe the rifins, the second family of clonally variant antigens known to be displayed by P. falciparum on the surface of the infected erythrocyte.

  15. 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

  16. 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.

  17. Typhoid fever in a Tertiary Hospital in Nigeria: Another look at the Widal agglutination test as a preferred option for diagnosis

    PubMed Central

    Enabulele, Osahon; Awunor, Simeon Nyemike

    2016-01-01

    Background: Single Widal agglutination test rather than blood culture, is commonly employed to diagnose typhoid fever in Nigeria. We took another look at the Widal agglutination test as a preferred option for diagnosis of typhoid fever by determining the specificity and sensitivity of Widal agglutination test in febrile adult patients. Materials and Methods: Two hundred and seventy-one blood samples from consecutive adults (>18 years) with febrile illness attending the General Practice Clinic of the University of Benin Teaching Hospital were tested using the Widal agglutination test, blood culture, and malaria parasite test on each sample to establish the diagnosis of typhoid fever. Results: Of the 271 blood samples 124 (45.76%) were positive following a Widal agglutination test, 60 (22.10%) blood samples grew Salmonella organisms on blood culture while 55 (20.29%) blood samples showed a co-infection of typhoid fever and malaria. A sensitivity of 35%, specificity of 51%, positive predictive value of 17%, and a negative predictive value of 73% were observed for Widal agglutination test as a diagnostic modality for typhoid fever infection. Conclusion: A single Widal agglutination test is not a valid diagnostic option for typhoid fever while co-infection with malaria parasite is the preponderant microbiological finding in typhoid fever infections. The severity of malaria parasitemia is associated with positive titers on Widal test. PMID:27397952

  18. Methane-induced haemolysis of human erythrocytes.

    PubMed Central

    Batliwala, H; Somasundaram, T; Uzgiris, E E; Makowski, L

    1995-01-01

    Human erythrocytes were exposed to high concentrations of methane and nitrogen through the application of elevated partial pressures of these gas molecules. Cell leakage (haemolysis) was measured for cells exposed to these gases under a wide range of experimental conditions. Application of methane produces haemolysis at pressures far below the hydrostatic pressures known to disrupt membrane or protein structure. The effects of changes in buffer, temperature, diffusion rate and detergents were studied. Methane acts co-operatively with detergents to produce haemolysis at much lower detergent concentration than is required in the absence of methane or in the presence of nitrogen. At sufficiently high concentrations of methane, all cells are haemolysed. Increased temperature enhances the effect. Methane produces 50% haemolysis at a concentration of about 0.33 M compared with about 7.5 M methanol required for the same degree of haemolysis. Images Figure 1 PMID:7733880

  19. 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.

  20. 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.

  1. Structural effects of titanium citrate on the human erythrocyte membrane.

    PubMed

    Suwalsky, M; Villena, F; Norris, B; Soto, M A; Sotomayor, C P; Messori, L; Zatta, P

    2005-03-01

    The structural effects of titanium citrate on the human erythrocyte membrane were studied through its interaction with intact erythrocytes and isolated unsealed human erythrocyte membranes (IUM). The studies were carried out by scanning electron microscopy and fluorescence spectroscopy, respectively. Titanium citrate induced shape changes in erythrocytes, which were damaged and ruptured leaving empty and retracted membranes. Fluorescence spectroscopy measurements in IUM indicated a disordering effect at both the polar head group and the acyl chain packing arrangements of the membrane phospholipid bilayer. Titanium citrate also interacted with molecular models of the erythrocyte membrane consisting in bilayers of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), representing classes of phospholipids located in the outer and inner monolayers of the erythrocyte membrane, respectively. X-ray diffraction indicated that titanium citrate induced structural perturbation of the polar head group and of the hydrophobic acyl regions of DMPC, while the effects on DMPE bilayers were negligible. This conclusion is supported by fluorescence spectroscopy measurements on DMPC large unilamellar vesicles. All these findings indicate that the structural perturbations induced by titanium to human erythrocytes can be extended to other cells, thereby affecting their functions. PMID:15708797

  2. 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.

  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. Participation of free oxygen radicals in damage of porcine erythrocytes

    SciTech Connect

    Jozwiak, J.; Helszer, Z.

    1981-10-01

    Gamma radiation causes disturbances in energy metabolism, decreases in (Na/sup +/-K/sup +/)-ATPase, Mg/sup 2 +/-APTase activity, and increase in the degree of hemolysis in porcine erythrocytes. Our results indicated a contribution of exogenous free radicals in radiation damage to porcine erythrocytes. In the presence of biological and chemical radioprotectors a protective effect with respect to ATPase activity and energy metabolism was observed in the presence of catalase, histidine, glucose, and sulfhydryl compounds. It appears that radiation damage to porcine erythrocytes is due to the action of various radicals formed upon irradiation which react at different rates with various cell constituents.

  5. 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.

  6. 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.

  7. Development of a Specific Latex Agglutination Test to Detect Antibodies of Enterovirus 71.

    PubMed

    Qin, Bo; Zhang, Jianhua; Xie, Wenhao; Liu, Xuehong; He, Tingting; Chen, Jinkun; Dong, Xuejun

    2015-10-01

    A latex agglutination test (LAT) was developed for the rapid detection of antibodies against the VP1 or VP1 proteins of Enterovirus 71 (EV71). The proteins of interest including prokaryotically expressed VP1 and two strains of anti-VP1 monoclonal antibody (McAb) against EV71 were covalently linked to carboxylated latex using ethyl-dimethyl-amino-propyl carbodiimide (EDC) to prepare sensitized latex beads. LAT was evaluated by an enzyme-linked immunosorbent assay (ELISA) as a reference test. The VP1-LAT showed a sensitivity of 87.0%, specificity of 88.9%, and an agreement ratio of 90.0% in detecting VP1 in 100 serum samples from experimentally infected mice, whereas these values were 86.8, 96.7, and 93.3%, respectively, for 608 clinical human serum samples. The VP1-LAT has advantages over other assays in terms of low cost, rapidity, chemical stability, high sensitivity, repeatability, and specificity. The LAT established in the present study is a rapid and simple test suitable for field monitoring of antibodies against VP1-EV71. PMID:26363276

  8. Comparison of fluorescent antibody and microscopic agglutination testing for Leptospira in pregnant and nonpregnant cows.

    PubMed

    Rajeev, Sreekumari; Berghaus, Roy D; Overton, Michael W; Pence, Mel E; Baldwin, Charles A

    2010-01-01

    Serum and urine samples from 30 cows (15 pregnant and 15 nonpregnant) from each of 10 Georgia dairy herds (total cows = 300) were examined by microscopic agglutination testing (MAT) and direct fluorescent antibody testing (FAT), respectively. Seven of the 10 herds had at least 1 cow with a positive FAT, and all of the herds had at least 1 cow with a reciprocal MAT titer > or =100 for 1 or more serovars. Serological testing was not helpful in identifying the infecting serovar for cows with a positive FAT result. The MAT titers for all 7 of the serovars evaluated were significantly correlated with one another, with 17 (81%) of the 21 Spearman rank correlation coefficients > or =0.4 in magnitude. Twenty (56%) of 36 FAT-positive cows did not have a titer that was highest for any particular serovar. Four of the 7 herds that reported using a Leptospira borgpetersenii serovar Hardjo-bovis vaccine had one or more FAT-positive cows compared with 3 out of 3 herds that reported they were not using this type of vaccine, although this difference was not statistically significant.

  9. Evaluation of a latex agglutination test (PYLOGEN) for the detection of Helicobacter pylori in stool specimens.

    PubMed

    Blanco, Silvia; Forné, Montse; Lacoma, Alicia; Prat, Cristina; Cuesta, Miguel Angel; Fuenzalida, Loreto; Viver, Josep Maria; Domínguez, Jose

    2009-04-01

    The aim of the study was to assess a new latex agglutination (LA) stool antigen assay (PYLOGEN; CerTest Biotec, Zaragoza, Spain) in the diagnosis of Helicobacter pylori infection and to monitor its eradication after treatment. The LA test has been approved for sale in Europe, and its approval from the US Food and Drug Administration is still pending. The individuals enrolled were classified into 3 groups of patients: Group 1 consisted of 38 patients who are H. pylori positive. The diagnosis of H. pylori infection was established if there was concordance between 2 test results (urea breath test [UBT], rapid urease test, and histopathologic study) or if the culture alone was positive. Patients with only 1 positive test were considered indeterminate and were excluded from the study. Group 2 comprised 9 patients without positive tests and who were considered to be H. pylori negative. Group 3 consisted of 57 patients who received eradication treatment. The sensitivity and specificity of the test were 78.9% and 100%, respectively. The results of the UBT of the patients were studied 6 weeks after eradication therapy. The sensitivity and specificity of the LA test relative to UBT for patients after treatment were 75% and 93.3%, respectively.

  10. The evolution of pretransfusion testing: from agglutination to solid-phase red cell adherence tests.

    PubMed

    Plapp, F V; Sinor, L T; Rachel, J M

    1989-01-01

    Hospital transfusion services and blood centers still use manual hemagglutination tests for most of their serological procedures. Automation of hemagglutination reactions has proven to be difficult, primarily because hemagglutination lacks an objective endpoint which can be easily interpreted by inexpensive instruments. Alternatively, solid-phase red cell adherence assays for ABO cell and serum grouping, Rh typing, red cell and platelet antibody screening, red cell and platelet crossmatching, IgA deficiency screening, hepatitis B surface antigen, and HIV antibody screening have been developed. The performance of these assays compares favorably with current hemagglutination and enzyme immunoassay methods. All of these tests share a common objective endpoint of adherence or nonadherence of indicator red cells. This uniformity allows easy interpretation of results visually, spectrophotometrically, or by image analysis. The latter technique has the potential to revolutionize the reading and interpretation of all agglutination tests. Solid-phase red cell adherence tests in microplates are ideal for batch processing large numbers of specimens. However, adherence tests are not restricted to this format. Therefore, blood grouping dipsticks have been produced, which permit testing of individual blood samples even outside of the laboratory.

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

    PubMed Central

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

    1983-01-01

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

  12. Simple solutions to false results with plate/slide agglutination tests in diagnosis of infectious diseases of man and animals

    PubMed Central

    Saxena, Hari Mohan; Chothe, Shubhada; Kaur, Paviter

    2015-01-01

    We have developed a new Superagglutination test for serodiagnosis of infectious diseases. It differs from conventional plate/slide agglutination tests (PAT/SAT) by three additional steps: prior staining of serum antibody by adding a dye and addition of diluted biotinylated antiglobulin and avidin in sequence after mixing the antigen with the test serum. The new steps circumvent the problems of false positive and false negative results of PAT/SAT. In serodiagnosis of brucellosis, Superagglutination test had higher positive predictive value and specificity than Rose Bengal Plate Test (RBPT) and Standard Tube Agglutination Test (STAT) and higher negative predictive value and sensitivity than RBPT, STAT, ELISA and Complement Fixation Test (CFT).•Superagglutination is a simple, accurate and economic screening test for infections.•More specificity, sensitivity, positive & negative predictive value than RBPT, STAT.•More sensitivity, negative predictive value than ELISA and Complement Fixation Test. PMID:26844209

  13. Evaluation of latex agglutination tests for fibrin-fibrinogen degradation products in the forensic identification of menstrual blood.

    PubMed

    Akutsu, Tomoko; Watanabe, Ken; Motani, Hisako; Iwase, Hirotaro; Sakurada, Koichi

    2012-01-01

    The identification of menstrual blood is important when discriminating menstruation from vaginal trauma in sexual assault cases. The aim of this study was to evaluate two fibrin-fibrinogen degradation product (FDP)-latex agglutination test kits, FDPL® Test (FDP-L) and FDP Plasma "RD" (FDP-P), for their ability to forensically identify menstrual blood. Sensitivity and specificity of the two kits were compared for menstrual blood and various body fluids, and the sensitivity of the FDP-latex agglutination test kit was also compared with that of an immunochromatographic test for human hemoglobin. The robustness of the FDP-latex agglutination test was compared with that of gene expression analysis of menstrual blood specific markers. The FDP-L kit was more sensitive than the FDP-P kit, but it cross-reacted with peripheral bloodstains from healthy volunteers. The FDP-P kit was specific for menstrual blood, with the exception of postmortem blood samples, and was not affected by other body fluids. In an FDP-negative menstrual blood sample, the sensitivity of human hemoglobin detection was lower than for FDP-positive samples and peripheral blood stains, suggesting that determination of human hemoglobin could be useful in interpreting negative results in the FDP-latex agglutination test. In menstrual blood samples incubated in wet conditions, FDP was found to be a robust marker in the identification of menstrual blood compared with mRNA markers. FDP-P testing was shown to be a suitable and highly efficient rapid screening test for the laboratory identification of menstrual blood.

  14. [Evaluation of a latex agglutination assay method for the determination of plasmin/alpha 2 plasmin inhibitor complex].

    PubMed

    Murakami, F; Iijima, K; Nakamura, K; Ikawa, S

    1994-06-01

    We evaluated a latex agglutination assay method for concentration of plasmin/alpha 2 plasmin inhibitor complex (PPI) developed recently. The latex reagent consisted of two kinds of latex particles, one was coated with monoclonal antibody against plasmin (JIPPI-3) and another coated with monoclonal antibody against modified alpha 2 plasmin inhibitor (JIPPI-50). A correlation of concentrations of PPI between this method and ordinary EIA kit was very good (r = 0.969). Within-run precision of latex agglutination reagent also was good. The concentrations of PPI in plasmas of 40 in 43 normal subjects were 0-0.8 microgram/ml and others were 0.8-1.6 microgram/ml. Plasma levels of PPI were markedly elevated in patients with DIC. In addition, half of the patients with malignant tumors or liver diseases had increased levels of PPI. 16 of 32 cases with selected diseases (18 malignant tumors, 4 liver diseases, 2 infectious diseases, 2 cerebral contusions, 6 others) showed abnormal levels in PPI (> or = 0.8 microgram/ml) during several days preceding the elevation of FDP. It suggested that PPI could reflect fibrinolysis earlier than FDP. This latex agglutination assay is a simple and rapid method, and specific for the determination of PPI concentration as well as EIA method. We conclude that this assay method is very convenient for clinical use. PMID:8051804

  15. Evaluation of Polysaccharide-Based Latex Agglutination Assays for the Rapid Detection of Antibodies to Burkholderia pseudomallei

    PubMed Central

    Suttisunhakul, Vichaya; Chantratita, Narisara; Wikraiphat, Chanthiwa; Wuthiekanun, Vanaporn; Douglas, Zakiya; Day, Nicholas P. J.; Limmathurotsakul, Direk; Brett, Paul J.; Burtnick, Mary N.

    2015-01-01

    Melioidosis is a severe disease caused by the Gram-negative bacterium Burkholderia pseudomallei. Diagnosis of melioidosis currently relies on the isolation of B. pseudomallei from clinical samples, which can take several days. An indirect hemagglutination assay (IHA) is widely used for serodiagnosis, but it has a short shelf life, is poorly standardized, and requires a viable bacteria culture performed in a biosafety level 3 (BSL-3) laboratory. To improve the diagnostic methods, we have developed two rapid latex agglutination tests based on purified B. pseudomallei O-polysaccharide (OPS) and capsular polysaccharide (CPS) antigens. The immunodiagnostic potential of these tests was evaluated using serum from culture-confirmed melioidosis patients (N = 143) and healthy donors from either endemic (N = 199) or non-endemic areas (N = 90). The sensitivity of the OPS-based latex agglutination assay (OPS-latex; 84.4%) was significantly higher than both the CPS-latex (69.5%) (P < 0.001) and IHA (69.5%) (P = 0.001). When evaluated with Thai donor serum, the OPS-latex had comparable specificity (56.9%) to the CPS-latex (63.8%) (P = 0.053), but was significantly lower than the IHA (67.6%) (P = 0.002). In contrast, all tests with U.S. donor serum were highly specific (≥ 97.8%). These results suggest that polysaccharide-based latex agglutination assays may be useful for serodiagnosis of melioidosis in non-endemic areas. PMID:26123956

  16. Detection of heat-stable antigens of Campylobacter jejuni and C. coli by direct agglutination and passive hemagglutination.

    PubMed

    Oza, A N; Thwaites, R T; Wareing, D R A; Bolton, F J; Frost, J A

    2002-03-01

    The two serotyping schemes for the detection of heat-stable antigens of Campylobacter jejuni and Campylobacter coli use the same strains for antiserum production but differ in the detection systems used for identifying agglutination. The Penner method uses passive hemagglutination (PHA) while the Laboratory of Enteric Pathogens method uses the same antisera but in a whole-bacterial-cell direct agglutination (DA) protocol. C. jejuni produces a polysaccharide capsule, which is antigenic, and is the main component detected by the PHA method. The DA method will detect both capsule antigens and lipopolysaccharide (LPS) or lipooligosaccharide (LOS) surface antigens. Comparison of both methods by using a selection of isolates from human infection has shown a range of variation in agglutination specificity, reflecting the differences in antigens detected by the two methods. While 27.4% of the 416 C. jejuni isolates reacted with the antisera raised against the same type strains by either method, the majority showed a range of more complex relationships. None of the 37 C. coli isolates reacted with the same antiserum by both methods. Together the two schemes gave a total of 102 distinct combined serogroups for C. jejuni and 16 for C. coli. Thus, while some clonally related isolates share the same capsule and LOS or LPS antigens, other strains appear to have a common capsule antigen but differ in their LPS or LOS structures or vice versa.

  17. 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.

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

    SciTech Connect

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

    1988-08-01

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

  19. 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.

  20. Malaria: a tumour necrosis factor inhibitor from parasitized erythrocytes.

    PubMed Central

    Sheikh, N A; Caro, H N; Taverne, J; Playfair, J H; Rademacher, T W

    1996-01-01

    The excessive production of tumour necrosis factor (TNF) is associated with the pathology of blood-stage malaria and phosphatidylinositol-containing phospholipid antigens from parasitized erythrocytes stimulate its secretion by macrophages, thus acting as toxins. This brief report describes some properties of an inhibitor present in lysates from erythrocytes infected with malarial parasites that blocked the detection of recombinant TNF in an enzyme-linked immunosorbent assay and diminished or abolished the cytotoxicity of TNF. It was not found in control lysates of normal erythrocytes. Its addition to macrophage cultures stimulated by toxic malarial preparations or by bacterial lipopolysaccharide also blocked the detection of TNF. These findings may explain the contradictory results obtained from different assays for TNF, and emphasize the need for caution when interpreting the results of a single assay system. If released when parasitized erythrocytes rupture in vivo, the inhibitor could help protect both parasite and host from the damaging effects of TNF. PMID:8778034

  1. The anticancer drug cisplatin interacts with the human erythrocyte membrane.

    PubMed

    Suwalsky, M; Hernández, P; Villena, F; Sotomayor, C P

    2000-01-01

    Drugs which exert their effects by interacting with DNA cause structural and functional membrane alterations which may be essential for growth inhibition by these agents. This paper describes the interaction of cisplatin with the human erythrocyte membrane and models constituted by bilayers of dimyristoylphosphatidylethanolamine (DMPE) and diacylphosphatidylserine (DAPS), representative of phospholipid classes located in the inner monolayer of the erythrocyte membrane, and of dimyristoylphosphatidylcholine (DMPC), a class present in its outer monolayer. Cisplatin ability to perturb DMPE, DAPS and DMPC bilayer structures was determined by X-ray diffraction and fluorescence spectroscopy. Electron microscopy disclosed that human erythrocytes incubated with 35 microM cisplatin, which is its therapeutical concentration in serum, developed cup-shaped forms (stomatocytes). According to the bilayer couple hypothesis, this means that the drug is inserted into the inner monolayer of the erythrocyte membrane, a conclusion supported by the studies on model systems. PMID:10928560

  2. [Erythrocytes and microvascular tone during acute traumatic haemorrhagic shock].

    PubMed

    Morel, N; Biais, M; Delaunay, F; Dubuisson, V; Cassone, O; Siméon, F; Morel, O; Janvier, G

    2013-05-01

    Haemorrhagic shock remains a leading cause of death in trauma patients. The concept of haematologic damage control is gradually taking place in the management of traumatic haemorrhagic shock. It is based primarily on the early implementation of a quality blood transfusion involving erythrocytes, plasmas and platelets transfusion. Red blood cell transfusion is mainly supported by the oxygen carrier properties of erythrocytes. However, it appears that erythrocytes ability to modulate the bioavailability of nitric oxide (NO) plays a major role in capillary opening and perfusion. Erythrocytes are also actively involved in the processes of hemostasis and coagulation. In this context, it seems difficult to define a threshold of hemoglobin concentration to determine the implementation of a blood transfusion in traumatic haemorrhagic shock.

  3. Teratology studies in the rabbit.

    PubMed

    Allais, Linda; Reynaud, Lucie

    2013-01-01

    The rabbit is generally the non-rodent species or second species after the rat recommended by the regulatory authorities and is part of the package of regulatory reproductive studies for the detection of potential embryotoxic and/or teratogenic effects of pharmaceuticals, chemicals, food additives, and other compounds, including vaccines (see Chapters 1-7).Its availability, practicality in housing and in mating as well as its large size makes the rabbit the preferred choice as a non-rodent species. The study protocols are essentially similar to those established for the rat (Chapter 9), with some particularities. The study designs are well defined in guidelines and are relatively standardized between testing laboratories across the world.As for the rat, large litter sizes and extensive background data in the rabbit are valuable criteria for an optimal assessment of in utero development of the embryo or fetus and for the detection of potential external or internal fetal malformations.

  4. Differential Fault Analysis of Rabbit

    NASA Astrophysics Data System (ADS)

    Kircanski, Aleksandar; Youssef, Amr M.

    Rabbit is a high speed scalable stream cipher with 128-bit key and a 64-bit initialization vector. It has passed all three stages of the ECRYPT stream cipher project and is a member of eSTREAM software portfolio. In this paper, we present a practical fault analysis attack on Rabbit. The fault model in which we analyze the cipher is the one in which the attacker is assumed to be able to fault a random bit of the internal state of the cipher but cannot control the exact location of injected faults. Our attack requires around 128 - 256 faults, precomputed table of size 241.6 bytes and recovers the complete internal state of Rabbit in about 238 steps.

  5. 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

  6. European Rabbits as Reservoir for Coxiella burnetii.

    PubMed

    González-Barrio, David; Maio, Elisa; Vieira-Pinto, Madalena; Ruiz-Fons, Francisco

    2015-06-01

    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.

  7. European Rabbits as Reservoir for Coxiella burnetii.

    PubMed

    González-Barrio, David; Maio, Elisa; Vieira-Pinto, Madalena; Ruiz-Fons, Francisco

    2015-06-01

    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

  8. 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

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

    PubMed

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

    2015-10-14

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

  10. Low toxicity method of inhibiting sickling of sickle erythrocytes

    DOEpatents

    Packer, Lester; Bymun, Edwin N.

    1977-01-01

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

  11. Reduced parasitemia observed with erythrocytes containing inositol hexaphosphate.

    PubMed Central

    Mintzer, C L; Deloron, P; Rice-Ficht, A; Durica, D; Struck, D K; Roessner, C A; Nicolau, C; Ihler, G M

    1988-01-01

    Chemicals entrapped in erythrocytes by hypotonic hemolysis can be assessed for possible antiparasitic activity both in vivo and in vitro, regardless of whether they are able to diffuse into erythrocytes readily. Inositol hexaphosphate, a highly charged compound, produced a dramatic lowering of the percentage of cells infected by Babesia microti in vivo and both B. microti and Plasmodium falciparum in vitro. Several possible mechanisms for this observation are discussed. PMID:3364957

  12. 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

  13. Arsanilic acid toxicity in rabbits.

    PubMed

    Confer, A W; Ward, B C; Hines, F A

    1980-04-01

    Rations from several rabbitries experiencing increased mortality, weight loss and diminished reproduction were analyzed for arsanilic acid. Levels of less than 56 ppm of arsanilic acid were found. A 30 day trial was conducted where arsanilic acid was given in doses of 1.6-16.2 mg/day in water to weanling and adult rabbits. The higher doses induced diarrhea, terminal convulsions and death. Weight loss or reduced weight gains occurred in six of seven treated groups. No significant gross or microscopic lesions were observed. Chemical analysis demonstrated the presence of increased total hepatic arsenic levels in treated compared to control rabbits.

  14. Correlation-timing-based erythrocyte velocity measurement using CCD imagery

    NASA Astrophysics Data System (ADS)

    O'Reilly, William J.; Hudetz, Anthony

    2001-05-01

    An automated correlation method is introduced to estimate erythrocyte velocity component of erythrocyte flux within the cerebral capillary network. Erythrocyte flux, defined as the number of red blood cells passing through a plane orthogonal to the axis of erythrocyte flow in a vessel per unit time, is considered to be the closest index of capillary flow. Introduced previously is the two-point cross-correlation method, a method whereby a video photometric analyzer captures the voltage produced from two electronic windows placed over a vessel of interest. In our new method, instead of using electronic windows, we use a CCD array, focused on a two- dimensional projection of the three-dimensional capillary structure. Simulations of this method yields accurate velocity measurements at a measured cell intensity of .2 standard deviations above mean noise values or cell counts fewer than 30 cells per minute for image sequences of 180 frames captured over a time interval of three seconds. We conclude that with proper reduction in the measured standard deviation of noise and by increasing the percentage of fluorscently labeled erythrocytes injected into the rat, the correlation timing method of estimating erythrocyte velocity is an accurate substitute for hand-measured velocity calculation.

  15. Recombinant outer membrane protein C of Aeromonas hydrophila elicits mixed immune response and generates agglutinating antibodies.

    PubMed

    Yadav, Sunita Kumari; Meena, Jitendra Kumar; Sharma, Mahima; Dixit, Aparna

    2016-08-01

    Aeromonas hydrophila is a gram-negative fish pathogenic bacterium, also responsible for causing opportunistic pathological conditions in humans. It causes a number of diseases in fish due to which the fish industry incurs huge economic losses annually. Due to problems of antibiotic resistance, and the rapidity with which the infection spreads among fishes, vaccination remains the most effective strategy to combat this infection in fish populations. Among various virulence factors associated with bacterial virulence, outer membrane proteins have been widely evaluated for their vaccine potential owing to their surface exposure and related role in pathogenicity. In the present study, we have investigated the immunogenic potential of a non-specific porin, outer membrane protein C (OmpC) whose expression is regulated by the two-component regulatory system and plays a major role in the survival of A. hydrophila under different osmolaric conditions. The full-length gene (~1 kb) encoding OmpC of A. hydrophila was cloned, characterized and expressed in E. coli. High yield (~112 mg/L at shake flask level) of the recombinant OmpC (rOmpC) (~40 kDa) of A. hydrophila was obtained upon purification from inclusion bodies using Ni(2+)-NTA affinity chromatography. Immunization with purified rOmpC in murine model generated high endpoint (>1:40,000) titers. IgG isotyping, ELISA and ELISPOT assay indicated mixed immune response with a TH2 bias. Also, the anti-rOmpC antibodies were able to agglutinate A. hydrophila in vitro and exhibited specific cross-reactivity with different Aeromonas strains, which will facilitate easy detection of different Aeromonas isolates in infected samples. Taken together, these data clearly indicate that rOmpC could serve as an effective vaccine against different strains of Aeromonas, a highly heterogenous group of bacteria. PMID:27328672

  16. Seroprevalence of Mycoplasma gallisepticum antibody by ELISA and serum plate agglutination test of laying chicken

    PubMed Central

    Ali, Md. Zulfekar; Rahman, Md. Mostafizer; Sultana, Shirin

    2015-01-01

    Aim: Mycoplasma gallisepticum (MG) is important avian pathogens responsible for chronic respiratory diseases of chicken and turkeys, which result in large economic loss for the poultry industry. The objectives of this study were determination of seroprevalence of MG antibody of commercial layer chicken at laying period in selected areas of Bangladesh. Materials and Methods: A total of 563 blood samples were collected randomly from selected commercial layer chickens at laying period during the period from July to December, 2013. Indirect enzyme linked immunosorbent assay (iELISA) and serum plate agglutination (SPA) test were performed to detect the presence of antibodies against MG. Results: Of 563 samples, 64.47% and 56.13% showed an overall prevalence of MG antibodies in iELISA and SPA test respectively. Prevalence of MG was recorded the highest (69.63%) at 50-55 weeks of age compared with lowest (53.26%) at 56-61 weeks of age (p<0.05). Significant (p<0.05) effect of breed were observed in the seroprevalence of MG infection in layer birds in the present study. The overall, 68.77%, 63.74% and 59.37% prevalence were found respectively in sonali, ISA Brown and White leg horn. The prevalence of MG antibodies was the highest (70.13%) in December followed by November (68%), October (65.67%), August (63.46%), September (58.54%) and July (51.78%) month. The seroprevalence of MG antibodies was higher (69.63%) in most of the large flocks and lower (56.82%) in small flocks. Conclusion: Therefore, might be suggested that the commercial layer farms should be routinely checked to monitor MG infection and the reactor birds should be culled since MG organism has the potential to transmit vertically. The correlation between MG antibody in month and flock size was not significant (p=0.359 and p=0.868, respectively). PMID:27046987

  17. Exfoliative toxin detection using reversed passive latex agglutination: clinical and epidemiologic applications.

    PubMed Central

    Kawabata, A; Ichiyama, S; Iinuma, Y; Hasegawa, Y; Ohta, M; Shimokata, K

    1997-01-01

    A rapid and simple method for detecting exfoliative toxin serotypes A and B from clinical isolates has been developed as a test kit (EXT-RPLA; Denka Seiken Co. Ltd., Niigata, Japan). This method is based on reversed passive latex agglutination. The detection limit of the EXT-RPLA observed for purified exfoliative toxin serotypes A and B was 1 ng/ml. We evaluated the clinical and epidemiologic uses of the EXT-RPLA. A total of 381 isolates of Staphylococcus aureus, 292 from various clinical specimens and 89 from the skin of dermatologic patients, were studied. The EXT-RPLA detected 19 exfoliative toxin producers, including 16 serotype A producers and 3 serotype B producers, but no double producers. The sensitivity and specificity of the EXT-RPLA were confirmed by the newborn mouse bioassay and a PCR assay for the structural genes for exfoliative toxin serotypes A and B (eta and etb, respectively). The overall positivity rate of exfoliative toxin producers was 5.0% (19 of 381), including 16 serotype A isolates and 3 serotype B isolates. Of the 89 isolates from the skin of dermatologic patients, 12 (13.5%) were positive for exfoliative toxin production. Only 2 (1.3%) of the 153 methicillin-resistant S. aureus isolates produced exfoliative toxin, while 17 (7.5%) of the 228 methicillin-sensitive isolates produced exfoliative toxin. The EXT-RPLA assay is a simple and reliable method for detecting exfoliative toxin, and we recommend its use for the rapid diagnosis of staphylococcal scalded skin syndrome. We also recommend its use for detection of this syndrome so that effective control measures can be taken against the spread of this syndrome. PMID:9230367

  18. Association between human erythrocyte calmodulin and the cytoplasmic surface of human erythrocyte membranes.

    PubMed

    Agre, P; Gardner, K; Bennett, V

    1983-05-25

    This report describes Ca2+-dependent binding of 125I-labeled calmodulin (125I-CaM) to erythrocyte membranes and identification of two new CaM-binding proteins. Erythrocyte CaM labeled with 125I-Bolton Hunter reagent fully activated erythrocyte (Ca2+ + Mg2+)-ATPase. 125I-CaM bound to CaM depleted membranes in a Ca2+-dependent manner with a Ka of 6 x 10(-8) M Ca2+ and maximum binding at 4 x 10(-7) M Ca2+. Only the cytoplasmic surface of the membrane bound 125I-CaM. Binding was inhibited by unlabeled CaM and by trifluoperazine. Reduction of the free Ca2+ concentration or addition of trifluoperazine caused a slow reversal of binding. Nanomolar 125I-CaM required several hours to reach binding equilibrium, but the rate was much faster at higher concentrations. Scatchard plots of binding were curvilinear, and a class of high affinity sites was identified with a KD of 0.5 nM and estimated capacity of 400 sites per cell equivalent for inside-out vesicles (IOVs). The high affinity sites of IOVs most likely correspond to Ca2+ transporter since: (a) Ka of activation of (Ca2+ + Mg2+)-ATPase and KD for binding were nearly identical, and (b) partial digestion of IOVs with alpha-chymotrypsin produced activation of the (Ca2+ + Mg2+)-ATPase with loss of the high affinity sites. 125I-CaM bound in solution to a class of binding proteins (KD approximately 55 nM, 7.3 pmol per mg of ghost protein) which were extracted from ghosts by low ionic strength incubation. Soluble binding proteins were covalently cross-linked to 125I-CaM with Lomant's reagent, and 2 bands of 8,000 and 40,000 Mr (Mr of CaM subtracted) and spectrin dimer were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiography. The 8,000 and 40,000 Mr proteins represent a previously unrecognized class of CaM-binding sites which may mediate unexplained Ca2+-induced effects in the erythrocyte.

  19. Role of reaction resistance in limiting carbon monoxide uptake in rabbit lungs.

    PubMed

    Heller, H; Schuster, K

    1998-06-01

    The contribution of reaction resistance to overall resistance to pulmonary carbon monoxide (CO) uptake [DLCO/(ThetaCO . Vc), where DLCO is lung CO diffusing capacity, ThetaCO is CO uptake conductance of erythrocytes, and Vc is pulmonary capillary blood volume] was determined in 10 anesthetized, paralyzed, and artificially ventilated rabbits. On the basis of the classical double-reciprocal equation of F. G. W. Roughton and R. E. Forster (J. Appl. Physiol. 11: 290-302, 1957), DLCO/(ThetaCO . Vc) was obtained by solving the relation DLCO/(ThetaCO . Vc) = 1 - 2/(DLNO/DLCO), where DLNO/DLCO represents the ratio between the respective single-breath diffusing capacities (DL) of nitric oxide (NO) and CO pulmonary capillary blood. The lungs of eight rabbits were inflated, starting from residual volume, by using 55 ml of indicator gas mixture (0.2% CO and 0.05% NO in nitrogen). DL values were calculated by taking the end-tidal partial pressures of CO and NO as analyzed by using a respiratory mass spectrometer. The overall value was DLCO/(ThetaCO . Vc) = 0.4 +/- 0.025 (mean +/- SD). Because of the use of O2-free indicator gas mixtures, the end-tidal O2 partial pressures were approximately 21 Torr. In one other rabbit, the application of 0.2% CO and 0.001% NO yielded DLCO/(ThetaCO . Vc) = 0.39; in the tenth rabbit, however, inspiratory volume was varied, and an identical value was found at functional residual capacity. We conclude that the contribution of reaction resistance to overall resistance to pulmonary CO uptake is independent of the inspiratory NO concentration used, including, with respect to the pertinent literature, the conclusion that in rabbits, dogs, and humans this contribution amounts to 40% when determined at functional residual capacity.

  20. 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

  1. Immunity to erythrocytic stages of malarial parasites.

    PubMed

    Long, C A; Daly, T M; Kima, P; Srivastava, I

    1994-01-01

    In those individuals who live in endemic areas, immunity to malaria is slow to develop and stage-specific. The nature and antigenic specificity of this response, which may involve components of both cell-mediated and humoral immunity, is not well understood. Rodent models provide useful systems to explore the spectrum of host responses that may contribute to resolution of erythrocytic-stage infection or possibly to pathogenesis. Moreover, these models allow identification of plasmodial molecules that can induce different types of host responses. Two different mouse model systems, Plasmodium yoelii yoelii and P. chabaudi adami are presented. These have been selected because resolution of infection by P. yoelii yoelii has been shown to require B cell-dependent mechanisms, while control of acute P. chabaudi adami infection can be achieved by T cell-dependent mechanisms. A monoclonal antibody that provides passive protection to P. yoelii challenge infection has been shown to recognize the cysteine-rich, carboxyl-terminal region of the merozoite surface protein-1. This region, obtained in an appropriate configuration from recombinant Escherichia coli, can induce significant protective immune responses in naive mice. In contrast, cell-mediated immune mechanisms make a major contribution to resolution of asexual-stage P. chabaudi adami infection. An empirical approach using continuous flow electrophoresis has identified several low molecular weight plasmodial proteins that can induce partial protective responses in susceptible hosts. These observations are briefly discussed with respect to human malaria.

  2. Erythrocyte exchange and leukapheresis in pregnancy.

    PubMed

    Perseghin, Paolo

    2015-12-01

    Hematological diseases in pregnancy should be carefully managed with a multidisciplinary approach, which should include obstetrics, hematology and, in selected patients, apheresis professionals. Hematological malignancies in pregnant women are rare, but the attending physicians should be aware that the use of cytotoxic drugs, tyrosine-kinase inhibitors or differentiating agents such as all-trans retinoic acid (ATRA) during the first trimester of pregnancy might be teratogenic and, in turn, induce fetal abnormalities or abortion. Thus, in pregnant patients with either acute or chronic leukemia presenting with symptomatic hyperleukocytosis, leukocytapheresis (LA) could be considered as a bridge therapeutic option. Furthermore, sickle cell disease (SCD) in pregnant women is usually managed only with supportive care, i.e. packed red blood cell (RBC) transfusion to prevent excessive hemoglobin decrease, hydration and prevention of acute sickling crisis. Nevertheless, selected patients at high risk for placental detachment due to vasoocclusive acute crisis or with multiple pregnancies may benefit from prophylactic erythrocyte exchange (EEX). Both LA and EEX must be carried out by well trained personnel and the patients (and the fetus) must be under close clinical and instrumental monitoring. In the present paper, recent indications for performing either LA or EEX in pregnant patients are reviewed. PMID:26621538

  3. Nuclear abnormalities of marine fish erythrocytes.

    PubMed

    Strunjak-Perovic, I; Topic Popovic, N; Coz-Rakovac, R; Jadan, M

    2009-07-01

    The aim of this study was to monitor erythrocyte nuclear abnormalities (NA) including micronuclei (MN) in cultured and wild sea bass Dicentrarchus labrax and wild mullet Mugil spp. Seasonal sampling was performed at seven locations along the eastern coast of the Adriatic Sea. The frequency of NA and MN was positively correlated to temperature (NA: P < 0.05, r = 0.11; MN P < 0.05, r = 0.10), and there was also a positive correlation between NA and MN frequency (P < 0.001, r = 0.43). The lowest NA and MN values for both fish species were recorded in spring, while the highest were recorded in autumn. Significantly higher frequency of NA was seen in D. labrax compared to Mugil spp., while MN frequency was low in both species and not significantly different. There was no significant difference in NA and MN frequency between cultured and wild D. labrax sampled in the same month, and there was no difference between wild Mugil spp. sampled near or far from fish farms. In view of sampling sites, the highest values were detected in fishes from the Limski Channel, the lowest from the Janjina location.

  4. Energized endocytosis in human erythrocyte ghosts.

    PubMed Central

    Schriei, S L; Bensch, K G; Johnson, M; Junga, I

    1975-01-01

    The mechanism of endocytosis in resealed human erythrocyte ghosts was studied. The energy for endocytosis or micropinocytosis appears to be derived from Mg-ATP, and membrane internalization is preceded by activation of a membrane-associated Ca,Mg-ATPase and by the active efflux of Ca. Endocytosis, Ca,Mg-ATPase activity, and active Ca efflux all require the presence of Mg. Furthermore, these three phenomena, endocytosis, Ca,Mg-ATPase activity, and active Ca extrusion, all have a concentration dependence on Ca such that low concentrations stimulate and higher concentrations inhibit the phenomena. The optimal concentration of Ca is identical for endocytosis, active Ca efflux, and Ca,Mg-ATPase. Morphologic studies indicated that while active Ca efflux and activation of the Ca,Mg-ATPase activity occurred promptly upon onset of incubation, there was a significant time delay before endocytosis occurred, which suggests that endocytosis additionally involved a more slowly functioning mechanicochemical mechanism. Ruthenium red, a specific inhibitor of Ca,Mg-ATPase and Ca transport, inhibited endocytosis in a concentration-related manner. Prostaglandins E1 and E2 had no measurable effect on ghost endocytosis, active Ca efflux, or Ca,Mg-ATPase activity. Images PMID:124748

  5. Red and black rice decrease atherosclerotic plaque formation and increase antioxidant status in rabbits.

    PubMed

    Ling, W H; Cheng, Q X; Ma, J; Wang, T

    2001-05-01

    The influence of white, red and black rice consumption on atherosclerotic plaque formation induced by hypercholesterolemia was investigated in rabbits. Male rabbits (n = 36) were divided into five groups. They were fed a normal laboratory purified diet (normal group, n = 6), a high cholesterol (0.5 g/100 g) diet (HC group, n = 6), a high cholesterol diet with 30 g/100 g white rice (WR group, n = 8), 30 g/100 g red rice (RR group, n = 8), or 30 g/100 g black rice (BR group, n = 8) for 10 wk. Blood samples were collected for lipid measurements and aorta were removed for assessment of atherosclerotic plaques at the end of the protocol. The oxidant and antioxidant status of blood, erythrocytes, liver and aorta was evaluated. The area of atherosclerotic plaque was 50% lower in rabbits fed the red or black rice diets than in those fed the white rice diet. Compared with the HC and WR groups, serum HDL cholesterol and apolipoprotein (apo) A-I concentration were greater (P < 0.05) in the RR and BR groups. Liver reactive oxygen species (ROS) and aortic malondialdehyde (MDA) were significantly lower, and the liver total antioxidative capacity (TAC) and erythrocyte superoxide dismutase (SOD) activity were significantly higher in the RR and BR groups compared with the HC and WR groups. Red or black rice consumption reduced or retarded the progression of atherosclerotic plaque development induced by dietary cholesterol. The enhanced serum HDL cholesterol and apo A-I concentrations, and the increased antioxidant and decreased oxidative status may be mechanisms of the antiatherogenic effect of red or black rice. PMID:11340093

  6. 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.

  7. 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

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

    PubMed Central

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

    2006-01-01

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

  9. Carbonic anhydrase isozymes of osteoclasts and erythrocytes of osteopetrotic microphthalmic mice.

    PubMed

    Jilka, R L; Rogers, J I; Khalifah, R G; Vaananen, H K

    1985-01-01

    The content of carbonic anhydrase isozymes I (CA I) and II (CA II) in red blood cells and bone of osteopetrotic microphthalmic (mi/mi) mice was analyzed. Monospecific rabbit polyclonal antibodies against purified rat carbonic anhydrase I or II detected both isozymes in hemolysates of both normal and mi/mi mice. Total carbonic anhydrase activity measurements of hemolysates from normal or mi/mi mice were identical. A procedure based on bromopyruvate inactivation was devised to measure the relative contributions of CA I and II isozymes to the carbon dioxide hydration activity of hemolyzates. CA II dominates the observed activity of hemolysates of normal and mi/mi mice. Immunohistochemical studies showed that CA II was present in osteoclasts of tibial and calvarial bones of both normal and mi/mi mice. Thus, in contrast to the several cases of inherited osteopetrosis in humans, there is no lack of active CA II in erythrocytes of mi/mi mice. The absence of CA II, therefore, does not appear to play a role in the etiology of osteopetrosis in the mi/mi mouse.

  10. 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.

  11. Antileukocyte antibody in postpartum and renal transplant subjects. A comparison of capillary agglutination and lymphocytotoxicity reactions.

    PubMed

    Thompson, J S; Jackson, D; Greazel, N A; Parmely, M J; Severson, C D

    1976-02-01

    Sera were obtained from 48 gravida II prenatal and 211 multiparous nonpregnant females and examined for leukocyte antibodies comparing a standard lymphocytotoxicity (CY) test with capillary agglutination (CA). Antibody was detected in 41% of the samples in both groups but only CA tests were positive with approximately one-half of the prenatal and three-fourths of the multiparous specimens. Although, CA reactions, when accompanied by positive CY responses, usually correlated with HLA, no correlation with HLA, 5b, or the neutrophil antigens was determined for 35 of the 48 sera reacting only by CA. As a model to test the specificity of CA positive-CY negative antisera, four extensively studied sera were further analyzed in 16 families. Independent segregation from the HLA complex and ABO and Rh antigens was confirmed and two of the sera appeared to detect separate clusters of reactions in conjunction with some of the other reagents. Pre- and postgraft samples obtained from 23 living related and 75 cadaveric renal transplanted patients were investigated and compared for graft function and prospective tissue typing. Although direct crossmatches were negative prior to surgery, 17.9% of the pretransplant samples from living related and 28.0% from cadaveric recipients contained detectable antibody when tested against a cell panel. Similar to the prenatal and multiparous groups, the majority of these responses were detected by CA. Following engraftment, antibody first became evident in 11 of 19 (58%) living related and in 23 of 53 (48.2%) cadaveric hosts. There was a striking association between the development of CA and CY antibody and failure, as contrasted to 100% 9-month or greater survival in 10 of 10 living related and 15 of 15 cadaveric transplants in whom only CA antibodies arose postoperatively. In total, these studies indicate that CA reacts with HLA antigens in common with CY tests. In addition, CA may detect HLA when CY is negative but many other reactions

  12. 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

  13. Evaluation of an Immunocapture-Agglutination Test (Brucellacapt) for Serodiagnosis of Human Brucellosis

    PubMed Central

    Orduña, Antonio; Almaraz, Ana; Prado, Ana; Gutierrez, M. Purificación; Garcia-Pascual, Agustina; Dueñas, Ana; Cuervo, Milagros; Abad, Ramon; Hernández, Beatriz; Lorenzo, Belen; Bratos, Miguel A.; Torres, Antonio Rodriguez

    2000-01-01

    We evaluated the validity and the usefulness of a new test for the diagnosis of human brucellosis based on an immunocapture-agglutination technique. A total of 315 sera from 82 patients with a diagnosis of brucellosis, 157 sera from patients in whom brucellosis was suspected but not confirmed, and 412 sera from people living in rural areas with endemic brucellosis were studied. The seroagglutination test (SAT), Coombs anti-Brucella test, and Brucellacapt test were evaluated. All the initial sera from the 82 patients proved to be positive in Brucellacapt and Coombs tests, while only 75 (91.4%) were positive in the SAT. If a ≥1/160 diagnostic threshold titer was defined for the Brucellacapt test, Coombs test, and SAT, the sensitivities were 95.1, 91.5, and 65.8%, respectively. Taking the same diagnostic threshold titer for the 157 sera from the unconfirmed but suspected patients, the specificities of the Brucellacapt, Coombs, and SAT were 81.5, 96.2, and 100%, respectively; for the 412 control sera, the specificities were 99.0, 99.8, and 100%. The diagnostic efficiency (area below the receiver operating characteristic curve) of Brucellacapt was 0.987852 (95% confidence interval [CI], 0.95109 to 0.99286), very similar to the diagnostic efficiency of the Coombs test (0.97611; 95% CI, 0.94781 to 0.99146) and higher than that of SAT (0.91013; 95% CI, 0.86649 to 0.94317). The results of the Brucellacapt test were compared with those of the Coombs test (correlation coefficient, 0.956; P = 0.000) and SAT (correlation coefficient, 0.866; P = 0.000). The study shows very good correlation between the Brucellacapt and Coombs tests, with a high concordance between titers obtained in the two tests. Nevertheless, lower correlation and concordance were found between the Brucellacapt and Coombs tests when the results for titers of ≥1/160 were compared (0.692; P = 0.000). In acute brucellosis, the Brucellacapt and Coombs tests render positive titers of ≥1/160. When the titers

  14. 9 CFR 354.124 - Quarantine of diseased rabbits.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Procedures; Ante-Mortem Inspections § 354.124 Quarantine of diseased rabbits. If live rabbits, which are... further handling of the rabbits will not create a health hazard, the lot shall be subject to ante-mortem... circumstances are such that release for treatment is impracticable, a careful rabbit-by-rabbit...

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

    PubMed

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

    2012-06-01

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

  16. Biochemistry of the erythrocyte Rh polypeptides: a review.

    PubMed Central

    Agre, P.; Smith, B. L.; Hartel-Schenk, S.

    1990-01-01

    The clinically important Rh blood group system is complex, consisting of multiple distinct antigens. Despite clinical recognition for over 50 years, the Rh blood group antigens have remained poorly understood on a molecular level until the recent identification and characterization of the "Rh polypeptides," the core structural proteins of the Rh antigens. This group of erythrocyte membrane proteins of molecular weight 30,000-35,000 daltons was first recognized by employing Rh-specific antibodies to immunoprecipitate radiolabeled components of erythrocyte membranes. By using antibodies specific for the Rh D, c, and E antigens, a series of highly related non-identical proteins were immunoprecipitated, indicating that the Rh antigens are composed of multiple related proteins. The Rh polypeptides have been purified and characterized, and they were found to have several unusual biochemical characteristics. The Rh polypeptides penetrate the membrane bilayer; they are linked to the underlying membrane skeleton; they are covalently fatty acid acylated with palmitate. While the Rh antigenic reactivity is unique to human erythrocytes, the Rh polypeptides have been isolated from erythrocytes of diverse species and are thought to be fundamental components of all mammalian erythrocyte membranes. The functional role of the Rh polypeptides remains undefined, but a role in the organization of membrane phospholipid is suspected. PMID:2127333

  17. Developmental patterns of antioxidant defense mechanisms in human erythrocytes.

    PubMed

    Ripalda, M J; Rudolph, N; Wong, S L

    1989-10-01

    To obtain a profile of erythrocyte antioxidant defense potential during late fetal development, we studied selected antioxidant parameters in blood samples from 65 neonates with birth wt between 520 and 4210 g and from 12 healthy adults. Erythrocyte superoxide dismutase activity did not change significantly with maturation and no significant differences were observed among preterm infants grouped in increasing birth wt categories, term neonates, and adults. Erythrocyte catalase and glutathione peroxidase, as well as plasma vitamin E levels, showed highly significant positive correlations (p less than 0.001) with increasing fetal wt and gestational age; by term, CAT activity reached a level similar to the adult control group, but glutathione peroxidase activity, as well as plasma vitamin E levels, were markedly lower in all the preterm and in the term groups than in adults (p less than 0.01). Erythrocyte glutathione S-transferase activity showed a negative correlation with increasing gestational age (p less than 0.01) and the adult values were considerably lower than any of the neonatal levels (p less than 0.001). The role of glutathione S-transferase in erythrocyte metabolism remains obscure. Maturational changes in the activity of the red cell enzymes that were studied and in the plasma vitamin E level were apparent from about 31-36 wk of gestation, suggesting that the stimulation for these changes may have commenced from about 28-31 wk.

  18. Effects of the antiepileptic drug carbamazepine on human erythrocytes.

    PubMed

    Suwalsky, Mario; Mennickent, Sigrid; Norris, Beryl; Villena, Fernando; Sotomayor, Carlos P

    2006-12-01

    The structural effects of the antiepileptic drug carbamazepine (CBZ) on the human erythrocyte membrane and molecular models have been investigated in the present work. This report presents the following evidence that CBZ interacts with red cell membranes: (a) X-ray diffraction and fluorescence spectroscopy of phospholipid bilayers showed that CBZ perturbed a class of lipids found in the outer moiety of the erythrocyte membrane; (b) in isolated unsealed human erythrocytes (IUM) the drug induced a disordering effect on the polar head groups and acyl chains of the membrane lipid bilayer; (c) in scanning electron microscopy (SEM) studies on human erythrocytes the formation of echinocytes was observed, due to the preferential insertion of CBZ in the outer monolayer of the red cell membrane. The effects of the drug detected in the present work were observed at concentrations of the order of those currently appearing in serum when it is therapeutically administered. This is the first time that toxic effects of carbamazepine on the human erythrocyte membrane have been described. PMID:16844339

  19. Morphological Effects Induced In Vitro by Propranolol on Human Erythrocytes.

    PubMed

    Suwalsky, Mario; Zambrano, Pablo; Villena, Fernando; Manrique-Moreno, Marcela; Gallardo, María José; Jemiola-Rzeminska, Malgorzata; Strzalka, Kazimierz; Edwards, Ana María; Mennickent, Sigrid; Dukes, Nathan

    2015-08-01

    Despite the extended use and well-documented information, there are insufficient reports concerning the effects of propranolol on the structure and functions of cell membranes, particularly those of human erythrocytes. Aimed to better understand the molecular mechanisms of its interactions with cell membranes, human erythrocyte and molecular models of the red cell membrane were utilized. The latter consisted of bilayers of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), representative of phospholipid classes located in the outer and inner monolayers of the human erythrocyte membrane, respectively. The capacity of propranolol to perturb the multibilayer structures of DMPC and DMPE was evaluated by X-ray diffraction. Moreover, we took advantage of the capability of differential scanning calorimetry to detect the changes in the thermotropic phase behavior of lipid bilayers resulting from propranolol interaction with DMPC and DMPE multilamellar vesicles. In an attempt to further elucidate their effects on cell membranes, the present work also examined their influence on the morphology of intact human erythrocytes by means of defocusing and scanning electron microscopy. Results indicated that propranolol induced morphological changes to human erythrocytes and interacted in a concentration-dependent manner with phospholipid bilayer. PMID:25724773

  20. 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

  1. Improvement of a latex agglutination test for the evaluation of oxacillin resistance in coagulase-negative staphylococci.

    PubMed

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

    2004-11-01

    The "Slidex MRSA Detection" test (Denka Seiken, Japan) is a latex agglutination assay able to detect PBP2a. We evaluated its ability to differentiate mecA-positive from mecA-negative coagulase-negative staphylococci. We included 100 coagulase-negative staphylococci clinical isolates belonging to 9 species, 54 mecA positive and 46 mecA negative, as characterized by PCR. The specificity achieved using the manufacturer's instructions was 100%, but the sensitivity was only 57%. To increase sensitivity, we introduced modifications into the standard protocol. Using either large inocula or oxacillin induction before test performance, we achieved 100% sensitivity.

  2. Differential agglutination by soybean agglutinin of human leukemia and neuroblastoma cell lines: potential application to autologous bone marrow transplantation.

    PubMed

    Reisner, Y

    1983-11-01

    Normal human bone marrow cells were mixed with radioactively labeled tumor cells from different leukemia and neuroblastoma cell lines, and the cell mixtures were separated by differential agglutination with soybean agglutinin. It is shown that the cell fraction unagglutinated by soybean agglutinin, which was previously found to be capable of reconstituting the hematopoietic system of lethally irradiated recipients, can be purged of tumor cells with varying efficiency depending on the tumor cell expression of soybean agglutinin receptors as detected by flow cytofluorimetry with fluoresceinated soybean agglutinin.

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

    PubMed Central

    2014-01-01

    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. PMID:24490832

  4. 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.

  5. [Effects of La3+ on calcium binding to erythrocyte cytoskeleton].

    PubMed

    Kravtsov, G M; Postnov, Iu V

    1992-02-01

    When the whole erythrocytes were exposed to LaCl3, A--23187, ionomycin, orthovanadate and saponin, there was Ca2+ binding only following La3+ treatment of the cells. The binding was evident at a wide range (0.1 microM--1.OmM) of La3+ concentrations. Iodoacetamide-induced (incubation for 3 hours, 37 degrees C) decrease in erythrocyte ATP levels was found to result in a 3-fold reduction in Ca2+ binding to the cytoskeleton. La(3+)-induced Ca2+ binding enhanced the incorporation of 14C-glucose and/or its metabolites into the red cell skeleton. Thus, the detected new type of Ca2+ binding to the cytoskeleton of human and rat erythrocytes is likely to be due to the cumulative process: direct binding of La3+ to the outer surface of a membrane and the metal-induced trigger of nucleotide--dependent intracellular process.

  6. 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

  7. 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.

  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. 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.

  10. [Structural-functional changes in erythrocyte membranes in bovine lympholeukemia].

    PubMed

    Riazantsev, V V; Kovalenko, L V; Belous, A M

    1996-01-01

    The condition of lipid peroxidation and activity of enzymes of protective glutathione-dependent anti-oxidation system of erythrocytes: glutathione peroxidase (GSH-P) and glutathione reductase (GSH-R) in cows with leukosis has been studied. The decrease of the level of MDA and GSH-R activity was accompanied by GSH-P activation depending on the stage disease. The considerable lowering of Ca2+ transport to erythrocytes was shown on hematological stage of leucosis. The qualitative composition of membrane proteins does not change according to gel electrophoresis data. But the quantity of main cytoskeleton protein, spectrin, increases in the "white shadows" of erythrocytes in the animals with leucosis. PMID:8755110

  11. 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.

  12. 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.

  13. 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.

  14. 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

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

    PubMed

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

    1999-01-01

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

  16. Rat Erythrocyte Insulin Receptors: Radioreceptor Assay and Characterization

    PubMed Central

    Ogunwole, John O.; Nerurkar, Shriniwas G.; Hollis, Vincent W.

    1985-01-01

    Highly specific insulin receptors have been identified on the rat erythrocyte. A radioreceptor assay for the evaluation of these receptors has been developed, and the characteristics of these receptors have been investigated. Insulin receptor binding on the rat erythrocytes was found to be dependent on pH, temperature, time, and ionic strength. When incubated for 3½ hours at 15° C, 5.0 × 109 erythrocytes/mL from each of 10 rats were found to bind specifically 7.54 percent (±0.15 SEM) of 40 pg of 125I-insulin. Specific binding was found to be a function of cell concentration. The pH optima for insulin binding were found to be 7.4 and 7.0 in the absence of cations. The presence of cations not only shifted pH optimum to 7.4 from 7.0, but also increased specific insulin binding. These observations suggest the stabilization of negatively charged groups on ligand and receptor, as well as providing a suitable ionic environment for the hormone-receptor interaction. Based on the resistance of rat erythrocytes to the pH of the external buffer, a simple method for determining the internal pH of rat red- blood cells is described. Scatchard analyses of insulin-binding data yielded curvilinear plots, and the number of receptor sites per cell was found to be 762 (±12.1 SD), as opposed to the large variation (410 ± 260 SD) in normal humans. The rat erythrocytes may serve as a useful, precise, sensitive, and efficient model system for future erythrocytic-receptor studies that would be difficult to obtain from human subjects. PMID:3981646

  17. Rat erythrocyte insulin receptors: radioreceptor assay and characterization.

    PubMed

    Ogunwole, J O; Nerurkar, S G; Hollis, V W

    1985-02-01

    Highly specific insulin receptors have been identified on the rat erythrocyte. A radioreceptor assay for the evaluation of these receptors has been developed, and the characteristics of these receptors have been investigated. Insulin receptor binding on the rat erythrocytes was found to be dependent on pH, temperature, time, and ionic strength. When incubated for 3½ hours at 15° C, 5.0 × 10(9) erythrocytes/mL from each of 10 rats were found to bind specifically 7.54 percent (±0.15 SEM) of 40 pg of (125)I-insulin. Specific binding was found to be a function of cell concentration. The pH optima for insulin binding were found to be 7.4 and 7.0 in the absence of cations. The presence of cations not only shifted pH optimum to 7.4 from 7.0, but also increased specific insulin binding.These observations suggest the stabilization of negatively charged groups on ligand and receptor, as well as providing a suitable ionic environment for the hormone-receptor interaction. Based on the resistance of rat erythrocytes to the pH of the external buffer, a simple method for determining the internal pH of rat red- blood cells is described. Scatchard analyses of insulin-binding data yielded curvilinear plots, and the number of receptor sites per cell was found to be 762 (±12.1 SD), as opposed to the large variation (410 ± 260 SD) in normal humans. The rat erythrocytes may serve as a useful, precise, sensitive, and efficient model system for future erythrocytic-receptor studies that would be difficult to obtain from human subjects.

  18. Erythrocyte stiffness during morphological remodeling induced by carbon ion radiation.

    PubMed

    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

  19. 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

  20. Cardiovascular physiology and diseases of the rabbit.

    PubMed

    Pariaut, Romain

    2009-01-01

    This article reviews what is known about the diagnosis and management of cardiovascular diseases in the pet rabbit. Current knowledge is based on anecdotal reports, derived from research data using the rabbit as an animal model of human cardiovascular diseases, but most importantly canine and feline cardiology. It is likely that, as cardiovascular diseases are more often recognized, more specific information will soon become available for the treatment of the pet rabbit with cardiac disease.

  1. [Sodium and calcium cation transport of erythrocytes in essential hypertension].

    PubMed

    Zhu, Z M; Song, K Q; Liu, G Y

    1989-08-01

    The sodium and calcium transport of erythrocyte and the influencing factors were studied in essential hypertensive (EH) subjects. The result showed that plasma endogenous digitalis-like compound (EDLC) increased and sodium pump depressed in some EH patients, but there were no parallel correlation between EDLC and sodium pump. The patients with normal sodium pump mainly showed their maximal Ca2+ pump activity and decreased calmodulin (CaM) content of erythrocyte. Thus there may be different types of ion transport defect in EH, and the abnormalities of these cation transports have an important role in the pathogenesis of EH. PMID:2560705

  2. Glycosylation of erythrocyte spectrin and its modification in visceral leishmaniasis.

    PubMed

    Samanta, Sajal; Dutta, Devawati; Ghoshal, Angana; Mukhopadhyay, Sumi; Saha, Bibhuti; Sundar, Shyam; Jarmalavicius, Saulius; Forgber, Michael; Mandal, Chhabinath; Walden, Peter; Mandal, Chitra

    2011-01-01

    Using a lectin, Achatinin-H, having preferential specificity for glycoproteins with terminal 9-O-acetyl sialic acid derivatives linked in α2-6 linkages to subterminal N-acetylgalactosamine, eight distinct disease-associated 9-O-acetylated sialoglycoproteins was purified from erythrocytes of visceral leishmaniaisis (VL) patients (RBC(VL)). Analyses of tryptic fragments by mass spectrometry led to the identification of two high-molecular weight 9-O-acetylated sialoglycoproteins as human erythrocytic α- and β-spectrin. Total spectrin purified from erythrocytes of VL patients (spectrin(VL)) was reactive with Achatinin-H. Interestingly, along with two high molecular weight bands corresponding to α- and β-spectrin another low molecular weight 60 kDa band was observed. Total spectrin was also purified from normal human erythrocytes (spectrin(N)) and insignificant binding with Achatinin-H was demonstrated. Additionally, this 60 kDa fragment was totally absent in spectrin(N). Although the presence of both N- and O-glycosylations was found both in spectrin(N) and spectrin(VL), enhanced sialylation was predominantly induced in spectrin(VL). Sialic acids accounted for approximately 1.25 kDa mass of the 60 kDa polypeptide. The demonstration of a few identified sialylated tryptic fragments of α- and β-spectrin(VL) confirmed the presence of terminal sialic acids. Molecular modelling studies of spectrin suggest that a sugar moiety can fit into the potential glycosylation sites. Interestingly, highly sialylated spectrin(VL) showed decreased binding with spectrin-depleted inside-out membrane vesicles of normal erythrocytes compared to spectrin(N) suggesting functional abnormality. Taken together this is the first report of glycosylated eythrocytic spectrin in normal erythrocytes and its enhanced sialylation in RBC(VL). The enhanced sialylation of this cytoskeleton protein is possibly related to the fragmentation of spectrin(VL) as evidenced by the presence of an additional 60

  3. Essential fructosuria: increased levels of fructose 3-phosphate in erythrocytes.

    PubMed

    Petersen, A; Steinmann, B; Gitzelmann, R

    1992-01-01

    Erythrocytes of 3 adult siblings with essential fructosuria contained 45-200 mumol/l fructose 3-phosphate (Fru-3-P), i.e. 3-15 times the concentration in normal controls. Sorbitol 3-phosphate was also increased, but to a lesser degree. An oral load with 50 g of fructose produced an additional 40 mumol/l increase of erythrocyte Fru-3-P after 5 h. The rate of Fru-3-P formation by red cells in vitro was normal. HbA1 and HbA1c were normal. The suspected pathogenetic role of Fru-3-P in diabetic complications is questioned.

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

    NASA Astrophysics Data System (ADS)

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

    2015-07-01

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

  5. Erythrocyte membrane fatty acids in multiple myeloma patients.

    PubMed

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

    2014-10-01

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

  6. Extraction of DNA from malaria-infected erythrocytes using isotachophoresis.

    PubMed

    Marshall, Lewis A; Han, Crystal M; Santiago, Juan G

    2011-12-15

    We demonstrate a technique for purification of nucleic acids from malaria parasites infecting human erythrocytes using isotachophoresis (ITP). We release nucleic acids from malaria-infected erythrocytes by lysing with heat and proteinase K for 10 min and immediately, thereafter, load sample onto a capillary device. We study the effect of temperature on lysis efficiency. We also implement pressure-driven counterflow during ITP extraction to extend focusing time and increase nucleic acid yield. We show that the purified genomic DNA samples are compatible with polymerase chain reaction (PCR) and demonstrate a clinically relevant limit of detection of 0.5 parasites per nanoliter using quantitative PCR.

  7. Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis

    PubMed Central

    Samanta, Sajal; Dutta, Devawati; Ghoshal, Angana; Mukhopadhyay, Sumi; Saha, Bibhuti; Sundar, Shyam; Jarmalavicius, Saulius; Forgber, Michael; Mandal, Chhabinath; Walden, Peter; Mandal, Chitra

    2011-01-01

    Using a lectin, Achatinin-H, having preferential specificity for glycoproteins with terminal 9-O-acetyl sialic acid derivatives linked in α2-6 linkages to subterminal N-acetylgalactosamine, eight distinct disease-associated 9-O-acetylated sialoglycoproteins was purified from erythrocytes of visceral leishmaniaisis (VL) patients (RBCVL). Analyses of tryptic fragments by mass spectrometry led to the identification of two high-molecular weight 9-O-acetylated sialoglycoproteins as human erythrocytic α- and β-spectrin. Total spectrin purified from erythrocytes of VL patients (spectrinVL) was reactive with Achatinin-H. Interestingly, along with two high molecular weight bands corresponding to α- and β-spectrin another low molecular weight 60 kDa band was observed. Total spectrin was also purified from normal human erythrocytes (spectrinN) and insignificant binding with Achatinin-H was demonstrated. Additionally, this 60 kDa fragment was totally absent in spectrinN. Although the presence of both N- and O-glycosylations was found both in spectrinN and spectrinVL, enhanced sialylation was predominantly induced in spectrinVL. Sialic acids accounted for approximately 1.25 kDa mass of the 60 kDa polypeptide. The demonstration of a few identified sialylated tryptic fragments of α- and β-spectrinVL confirmed the presence of terminal sialic acids. Molecular modelling studies of spectrin suggest that a sugar moiety can fit into the potential glycosylation sites. Interestingly, highly sialylated spectrinVL showed decreased binding with spectrin-depleted inside-out membrane vesicles of normal erythrocytes compared to spectrinN suggesting functional abnormality. Taken together this is the first report of glycosylated eythrocytic spectrin in normal erythrocytes and its enhanced sialylation in RBCVL. The enhanced sialylation of this cytoskeleton protein is possibly related to the fragmentation of spectrinVL as evidenced by the presence of an additional 60 kDa fragment, absent in

  8. Abnormalities in the erythrocyte membrane in acute lymphoid leukaemia.

    PubMed Central

    Kundu, M; Basu, J; Chakrabarti, P; Rakshit, M M

    1989-01-01

    Erythrocytes from patients suffering from acute lymphoid leukaemia (ALL) show decreased proportions of spectrin tetrameters and altered spatial distribution of band 4.1 and ankyrins. These abnormalities of the cytoskeleton are probably responsible for altered membrane fluidity and transbilayer distribution of phosphatidylethanolamine in ALL. ALL is associated with severe anaemia and usually, but not always, with overproduction of lymphocytes. To our knowledge, this is the first report of abnormalities in the erythrocyte membrane in ALL which may, in part, be responsible for the observed anaemia. PMID:2730573

  9. Comparison of three commercial rapid agglutination test kits for identification of coagulase positive staphylococci from foods and animals.

    PubMed

    Holme, I J; Rosef, O; Ewald, S

    1991-01-01

    Three rapid agglutination assays for the identification of Staphylococcus aureus Monostaph (Bionor A/S, Skien, Norway), Staphyslide-Test (BioMerieux, Lyon, France) and Staph-Rapid-Test (Roche, Basel, Switzerland), were compared. A total of 104 Gram-positive, catalase positive cocci were tested: Nineteen Staphylococcus reference strains comprising 15 spp. (4 strains were coagulase positive), and 7 Micrococcus reference strains comprising 4 spp.; 22 food isolates comprising 13 S. aureus, 8 coagulase positive Staphylococcus spp., and 1 Micrococcus sp.; 56 animal isolates comprising 11 S. aureus, 9 S. hyicus subsp. hyicus, 2 S. intermedius, 15 coagulase positive and 19 coagulase negative Staphylococcus spp. Totally 54 strains were coagulase positive. Considering agglutination of a coagulase positive strain as a correct identification, Monostaph, Staph-Rapid-Test, and Staphyslide-Test correctly identified 52 (96.3%), 47 (87.0%) and 48 (89.0%) of the coagulase positive staphylococci, respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test showed 1 (2.0%), 4 (8.0%) and 4 (8.0%) false positive reactions respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test gave 0 (0.0%), 6 (5.8%) and 7 (6.7%) non-interpretable reactions, respectively. Monostaph may be a good alternative to the tube-coagulase test for rapid and reliable identification of coagulase positive staphylococci from both food and veterinary sources. However, false negative reactions may occur with coagulase positive strains of S. hyicus subsp. hyicus and S. intermedius.

  10. Minimization of bacterial size allows for complement evasion and is overcome by the agglutinating effect of antibody

    PubMed Central

    Dalia, Ankur B.; Weiser, Jeffrey N.

    2011-01-01

    SUMMARY The complement system, which functions by lysing pathogens directly or by promoting their uptake by phagocytes, is critical for controlling many microbial infections. Here we show that in Streptococcus pneumoniae, increasing bacterial chain length sensitizes this pathogen to complement deposition and subsequent uptake by human neutrophils. Consistent with this, we show that minimizing chain length provides wild-type bacteria with a competitive advantage in vivo in a model of systemic infection. Investigating how the host overcomes this virulence strategy, we find that antibody promotes complement-dependent opsonophagocytic killing of Streptococcus pneumoniae and lysis of Haemophilus influenzae independent of Fc-mediated effector functions. Consistent with the agglutinating effect of antibody, F(ab′)2 but not Fab could promote this effect. Therefore, increasing pathogen size, whether by natural changes in cellular morphology or via antibody-mediated agglutination, promotes complement-dependent killing. These observations have broad implications for how cell size and morphology can affect virulence among pathogenic microbes. PMID:22100164

  11. [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.

  12. 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

  13. 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.

  14. 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.

  15. Monounsaturated fatty acid ether oligomers formed during heating of virgin olive oil show agglutination activity against human red blood cells.

    PubMed

    Patrikios, Ioannis S; Mavromoustakos, Thomas M

    2014-01-29

    The present work focuses on the characterization of molecules formed when virgin olive oil is heated at 130 °C for 24 h open in air, which are found to be strong agglutinins. The hemagglutinating activity of the newly formed molecule isolated from the heated virgin olive oil sample was estimated against human red blood cells (RBCs). Dimers and polymers (high molecular weight molecules) were identified through thin layer chromatography (TLC) of the oil mixture. (1)H and (13)C nuclear magnetic resonance (NMR) and gas chromatography-mass spectroscopy (GC-MS) were the methods used for structural characterization. Among others, oligomerization of at least two monounsaturated fatty acids (FA) by an ether linkage between the hydrocarbon chains is involved. Light microscopy was used to characterize and visualize the agglutination process. Agglutination without fusion or lysis was observed. It was concluded that the heating of virgin olive oil open in air, among other effects, produces oligomerization as well as polymerization of unsaturated FA, possibly of monohydroxy, monounsaturated FA that is associated with strong hemagglutinating activity against human RBCs. The nutritional value and the effects on human health of such oligomers are not discussed in the literature and remain to be investigated. PMID:24410166

  16. [Evaluation of commercial usefulness for microparticle agglutination Serodia-Myco II test for serodiagnosis of Mycoplasma pneumonia infections].

    PubMed

    Rastawicki, Waldemar; Kałuzewski, Stanisław; Jagielski, Marek; Gierczyński, Rafał

    2002-01-01

    The usefulness of Serodia-Myco II agglutination test (Fujirebio, Japan) for diagnosis of the M. pneumoniae infections was evaluated. A total of 66 serum samples obtained from patients with respiratory tract infections were tested by Serodia-Myco II test, complement fixation (CF) test, ELISA-IgG/-IgM, and by latex agglutination (LA) test prepared in our laboratory. Using CF test and ELISA as the reference tests, Serodia-Myco II test gave too many false positive results. This test in relation to CF test, ELISA-IgM, ELISA-IgG, and LA test showed a very high sensitivity, virtually 100%, with a low specificity, below 50%. It seems that oversensitivity of the Serodia-Myco II test is caused by too low cut off (40) value recommended by the manufacturer. The Serodia-Myco II test may be used in routine serodiagnosis of mycoplasmosis under condition that cut off value will be raised to 160 and the positive results of this test will be confirmed by the CF test or ELISA.

  17. Prevalence of agglutinating antibodies to Toxoplasma gondii in adult and fetal mule deer (Odocoileus hemionus) from Nebraska.

    PubMed

    Lindsay, David S; McKown, Richard D; DiCristina, Jennifer A; Jordan, Carly N; Mitchell, Sheila; Oates, David W; Sterner, Mauritz C

    2005-12-01

    Toxoplasma gondii is an apicomplexan parasite of mammals and birds. Herbivores acquire postnatal infection by ingesting oocysts from contaminated food or water. Toxoplasma gondii infection is common in white-tailed deer, Odocoileus virginianus, but little is known about the prevalence of infection in mule deer, O. hemionus. We examined sera from 89 mule deer from Nebraska for agglutinating antibodies to T. gondii using the modified direct agglutination test (MAT) with formalin-fixed tachyzoites as antigen. Thirty-one (35%) of the samples were positive at dilutions of > or = 1:25. Samples were examined from 29 fetuses from these mule deer and none were positive in the MAT. Sera from 14 white-tailed deer from Nebraska were also examined and 6 (43%) were positive for T. gondii. Samples were examined from 5 fetuses from these white-tailed deer and none was positive in the MAT. Our results in both deer species from Nebraska are similar to studies conducted in white-tailed deer from other regions of the United States. Our findings indicate that mule deer are frequently infected with T. gondii and that mule-deer meat may be a source of human infection.

  18. A comparison of erythrocyte glutathione S-transferase activity from human foetuses and adults.

    PubMed Central

    Strange, R C; Johnston, J D; Coghill, D R; Hume, R

    1980-01-01

    Glutathione S-transferase activity was measured in partially purified haemolysates of erythrocytes from human foetuses and adults. Enzyme activity was present in erythrocytes obtained between 12 and 40 weeks of gestation. The catalytic properties of the enzyme from foetal cells were similar to those of the enzyme from adult erythrocytes, indicating that probably only one form of the erythrocytes enzyme exists throughout foetal and adult life. PMID:7396875

  19. Study of the Structure, Oxygen-Transporting Functions, and Ionic Composition of Erythrocytes at Vascular Diseases

    PubMed Central

    Revin, Viktor V.; Gromova, Natalia V.; Revina, Elvira S.; Mel'nikova, Natalya A.; Balykova, Larisa A.; Solomadin, Ilia N.; Tychkov, Alexander Yu.; Revina, Nadezhda V.; Gromova, Oksana Yu.; Anashkina, Irina V.; Yakushkin, Viktor A.

    2015-01-01

    The present paper explores the role of erythrocytes in the pathogenesis of vascular diseases. The state of erythrocytes, their ionic composition and structure, and properties of erythrocytes hemoglobin were studied by using laser interference microscopy, Raman scattering spectroscopy, and capillary electrophoresis. In patients suffering from vascular disorders we identified statistically significant changes in the shape of erythrocytes, their ionic composition, and redistribution of hemoglobin throughout cells. PMID:26601112

  20. 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.

  1. Erythrocyte gangliosides act as receptors for Neisseria subflava: identification of the Sia-1 adhesin.

    PubMed Central

    Nyberg, G; Strömberg, N; Jonsson, A; Karlsson, K A; Normark, S

    1990-01-01

    Neisseria gonorrhoeae was recently shown to bind to a subset of lactose-containing glycolipids (N. Strömberg, C. Deal, G. Nyberg, S. Normark, M. So, and K.-A. Karlsson, Proc. Natl. Acad. Sci. USA 85:4902-4906, 1988). A number of commensal Neisseria strains were also shown to be lactose binders. In addition, Neisseria subflava bound to immobilized gangliosides, such as hematoside and sialosyl paragloboside, carrying the NeuAc alpha 2-3Gal beta 1-4Glc sequence. To a lesser extent, N. gonorrhoeae also bound to this receptor in vitro. In N. subflava GN01, this binding property mediated agglutination of human erythrocytes in a neuraminidase-sensitive fashion. Nitrosoguanidine-induced nonhemagglutinative mutants of N. subflava GN01 had lost the ability to bind hematoside and sialosylparagloboside but remained able to bind lactosylceramide and gangliotetraosylceramide. These mutants fell into three classes with respect to their outer membrane protein profiles in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Class 1 mutants were identical to the parent strain save for the loss of a 27-kilodalton (kDa) protein. Class 2 mutants showed an outer membrane protein profile identical to that of the wild type, whereas mutants belonging to class 3 showed a number of changes, including the apparent absence of the 27-kDa protein. The 27-kDa protein from N. subflava GN01 was purified from the supernatant. A polyclonal antiserum to the purified Sia-1 protein as well as a Sia-1-specific monoclonal antibody inhibited hemagglutination by strain GN01. The purified Sia-1 protein in the presence of diluted anti-Sia-1 antiserum mediated a neuraminidase-sensitive hemagglutination. The purified Sia protein from a class 2 mutant was not able to hemagglutinate when cross-linked with antibodies, suggesting that it is a mutant form of Sia-1 affected in the receptor-binding site. Immunoelectron microscopy with a Sia-1-specific monoclonal antibody revealed that the adhesin was

  2. 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.

  3. Adherence characteristics of attaching and effacing strains of Escherichia coli from rabbits.

    PubMed Central

    Robins-Browne, R M; Tokhi, A M; Adams, L M; Bennett-Wood, V; Moisidis, A V; Krejany, E O; O'Gorman, L E

    1994-01-01

    Twelve strains of Escherichia coli previously reported to cause diarrhea in rabbits were examined for properties associated with virulence. Ten strains met the criteria for classification as enteropathogenic E. coli in that they were diarrheagenic strains that evoked attaching-effacing lesions in the small intestine and did not produce detectable enterotoxins or cytotoxins. These bacteria exhibited a variety of patterns when investigated for adherence to HEp-2 epithelial cells. Although several strains displayed localized and/or diffuse adherence to epithelial cells, they did not hybridize with DNA probes that recognize the genes responsible for these phenotypes in diarrheagenic E. coli from humans. The bacteria also varied in their ability to bind to erythrocytes and intestinal brush borders from various animal species. Six strains adhered to rabbit brush borders; two of these also adhered to brush borders from other animals. Two strains that did not adhere to rabbit brush borders adhered to those from guinea pigs or sheep. Only one of the strains investigated carried AF/R1 fimbriae, which are believed to govern the host specificity of this category of diarrheagenic E. coli. This strain was E. coli RDEC-1, which remains the only E. coli strain to date that is known to carry fimbriae of this type. The results indicate that although diarrheagenic E. coli strains from rabbits may have common properties associated with the ability to produce attaching-effacing lesions, they differ from each other and from enteropathogenic E. coli of humans in terms of some of the adhesins that mediate binding to eukaryotic cells. Images PMID:8168918

  4. Comparison of sensitivities and specificities of latex agglutination and an enzyme-linked immunosorbent assay for detection of antibodies to the human immunodeficiency virus in African sera.

    PubMed

    Francis, H L; Kabeya, M; Kafuama, N; Riggins, C; Colebunders, R; Ryder, R; Curran, J; Izaley, L; Quinn, T C

    1988-11-01

    The sensitivities, specificities, and positive and negative predictive values of the Cambridge BioScience Corp. (Worcester, Mass.) human immunodeficiency virus latex agglutination assay were compared by using three different blood preparations. By using the manufacturer's standard test method with diluted sera, the sensitivity of latex agglutination was 100%, the specificity was 99.58%, and the positive and negative predictive values were 99.26 and 100%, respectively. Use of diluted whole blood or undiluted whole blood did not significantly affect the sensitivity (mean, 99.72%), specificity (mean, 99.47%), positive predictive value (mean, 99.07%), or negative predictive value (mean, 99.89%). The latex agglutination assay is a simple, rapid assay for the detection of human immunodeficiency virus that would be useful in Third World countries or other areas where enzyme-linked immunosorbent assays are not available or cannot be used.

  5. 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.

  6. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme...

  7. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme...

  8. 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...

  9. 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...

  10. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase... § 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay. (a) Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme...

  11. 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. PMID:7863738

  12. [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.

  13. In vivo biodistribution of a radiolabeled blood substitute: sup 99m Tc-labeled liposome-encapsulated hemoglobin in an anesthetized rabbit

    SciTech Connect

    Rudolph, A.S.; Goins, B. ); Klipper, R.W.; Phillips, W.T. )

    1991-12-01

    Liposome-encapsulated hemoglobin (LEH) is an erythrocyte substitute that is a potential resuscitative fluid for the in vivo delivery of oxygen. The authors had noninvasively imaged radiolabeled LEH in vivo with technetium-99m ({sup 99m}Tc) to study the biodistribution in an anesthetized rabbit. Rabbits were infused with 30 ml of LEH and imaged with a {gamma} camera continuously for 2 hr. At 20 hr postinfusion, the animals were imaged again and sacrificed; the organs were weighed and their radioactivity was determined for autopsy organ distribution. Organ uptake from the images was corrected for organ-associated blood pool, which was determined by infusion of {sup 99m}Tc-labeled rabbit erythrocytes. Blood pool and decay-corrected biodistribution data reveal the kinetics of LEH distribution. Image biodistribution data was also validated at 20 hr by tissue sampling. At 20 hr postinfusion, autopsy biodistribution data reveals approximately 42.6% of the total counts remaining in the blood, 15.4% in the liver, 18.1% in spleen, 3.2% in the lungs, 2.4% in muscle, 1.6% in urine, and trace levels in the kidney, brain, and heart (<1%). There is no evidence of hemoglobin release from LEH or kidney dysfunction at any time over the course of the study.

  14. 21 CFR 864.6700 - Erythrocyte sedimentation rate test.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

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

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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.

  20. The effect of bromfenvinphos and its impurities on human erythrocyte.

    PubMed

    Szatkowska, Bozena; Bukowska, Bozena; Huras, Bogumiła

    2011-02-01

    Bromfenvinphos - (E,Z)-O,O-diethyl-O-[1-(2,4-dichlorophenyl)-2-bromovinyl] phosphate (BFVF) is the insecticide elaborated in Poland, which has been used against Varroa destructor causing honey bees disease called as varroosis. The substances that are formed as a result of bromfenvinphos synthesis are dihydro-bromfenvinphos (O,O-diethyl O-[1-(2,4-dichlorophenyl)vinyl] phosphate); dibromo-bromfenvinphos (O,O-diethyl O-[1-(2,4-dichlorophenyl)-2,2-dibromovinyl] phosphate); 2,4-dichlorophenacyl bromide; 2,4-dichlorophenacylidene bromide and 2,4-dichlorophenacylidyne bromide. In this work, we evaluated the effect of these compounds on hemolysis and hemoglobin oxidation (met-Hb formation) in human erythrocytes. Moreover, the changes in the size (FSC-A) and the shape (SSC-A) of red blood cells were assessed using flow cytometry and phase contrast microscopy. It was proven that bromfenvinphos at concentrations ranging from 0.5 to 250 μM during 1h incubation did not change the parameters examined in human erythrocytes. Similarly, most of bromfenvinphos impurities did not increase hemolysis and methemoglobin level nor changed the size and shape of the erythrocytes. The exception was dibromo-bromfenvinphos, which changed the FSC-A and SSC-A parameters, as well as 2,4-dichlorophenacyl bromide which induced hemolysis, increased the level of met-Hb and changed erythrocytes morphology.

  1. 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...

  2. Age-related carbonyl stress and erythrocyte membrane protein carbonylation.

    PubMed

    Li, Guolin; Liu, Li; Hu, Hui; Zhao, Qiong; Xie, Fuxia; Chen, Keke; Liu, Shenglin; Chen, Yaqin; Shi, Wang; Yin, Dazhong

    2010-01-01

    Reactive carbonyl species (RCS) have been widely used as indicators of oxidative stress. However, the associations of carbonyl stress with aging process and biochemical alteration of erythrocyte are still poorly understood. Fresh blood samples in vacutainer tubes containing sodium heparinate were obtained from 874 volunteers who were divided into young, adult and old groups based on their age. Plasma RCS and thiols concentrations between different age groups and erythrocyte membrane protein carbonylation in the adult group were detected within 24h of the blood sampling. Results showed that the plasma thiols concentration decreased gradually during aging process, and the p-values between all three groups are less than 0.05. The plasma RCS concentration in different age groups showed a nonlinear association with age. The levels in the young group were slightly higher than the adult group (not significant) and lower than the old group (p < 0.01). The protein carbonylation of erythrocyte membrane was positively correlated with plasma RCS concentration (p < 0.01), but not plasma thiols concentration. We conclude that higher levels of RCS, not lower levels of thiols, in plasma are a direct risk factor for the protein carbonylation of erythrocyte membrane. Owing to the decrease of thiols levels and increase of RCS levels during aging process, a shift from RCS-related redox allostasis to carbonyl stress would contribute to age-related biological dysfunction and even aging process.

  3. The anticancer drug adriamycin interacts with the human erythrocyte membrane.

    PubMed

    Suwalsky, M; Hernández, P; Villena, F; Aguilar, F; Sotomayor, C P

    1999-01-01

    Adriamycin is an aminoglycosidic anthracycline antibiotic widely used in the treatment of cancer. Increasing reports point to the involvement of cell membranes in its mechanism of action. The interaction of adriamycin with human erythrocytes was investigated in order to determine the membrane binding sites and the resultant structural perturbation. Electron microscopy revealed that red cells incubated with the therapeutical concentration of the drug in human plasma changed their discoid shape to both stomatocytes and echinocytes. According to the bilayer couple hypothesis, this means that adriamycin was incorporated into either the inner or outer leaflets of the erythrocyte membrane. To explain this unusual result, the drug was incubated with molecular models. One of them consisted of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) multilayers, representative of phospholipid classes located in the outer and inner leaflets of the erythrocyte membrane, respectively. X-ray diffraction showed that adriamycin interaction perturbed the polar head and acyl chain regions of both lipids. Fluorescence spectroscopy on another model, consisting of DMPC large unilamellar vesicles (LUV), confirmed the X-ray results in that adriamycin fluidized its hydrophobic moiety. It is concluded that adriamycin incorporates into both erythrocyte leaflets affecting its membrane structure. PMID:10349743

  4. 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...

  5. [VISCOELASTISITY AND ELECTRICAL PARAMETERS OF ERYTHROCYTES IN GILBERT SYNDROME].

    PubMed

    Kurilovich, S A; Nemtsova, E G; Kruchinina, M V; Maximov, V N

    2015-01-01

    Results of viscoelastic and electrical properties of erythrocytes study in patients with genetically confirmed Gilbert's syndrome (n = 81) are presented. Dielectrophoresis of erythrocytes in a nonuniform an alterning electric field was performed in81 patients with Gilbert's syndrome and in 20 persons of the comparison group without of the pathology identified by thelaboratory and instrumental examination. The significant differences in viscoelasticity properties of erythrocytes in Gilbert'ssyndrome were obtained. The amplitude of the deformation, the speed of movement to the electrodes and the polarizability on electric field's of all frequencies were significantly lower, but generalized rigidity index, viscosity, index of aggregationand degradation on electric field's of all frequencies were higher than in the comparison group. A number of electricalparameters (conductivity, the capacity of the cells and the relative polarizability) were also higher than in the comparisongroup. Some differences in the parameters of erythrocytes were obtained from homozygous and heterozygous carriers of A(TA), TAA of gene UGT1A1 promotor. PMID:27214984

  6. 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.

  7. 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.

  8. Effect of high glucose concentrations on human erythrocytes in vitro

    PubMed Central

    Viskupicova, Jana; Blaskovic, Dusan; Galiniak, Sabina; Soszyński, Mirosław; Bartosz, Grzegorz; Horakova, Lubica; Sadowska-Bartosz, Izabela

    2015-01-01

    Exposure to high glucose concentrations in vitro is often employed as a model for understanding erythrocyte modifications in diabetes. However, effects of such experiments may be affected by glucose consumption during prolonged incubation and changes of cellular parameters conditioned by impaired energy balance. The aim of this study was to compare alterations in various red cell parameters in this type of experiment to differentiate between those affected by glycoxidation and those affected by energy imbalance. Erythrocytes were incubated with 5, 45 or 100 mM glucose for up to 72 h. High glucose concentrations intensified lipid peroxidation and loss of activities of erythrocyte enzymes (glutathione S-transferase and glutathione reductase). On the other hand, hemolysis, eryptosis, calcium accumulation, loss of glutathione and increase in the GSSG/GSH ratio were attenuated by high glucose apparently due to maintenance of energy supply to the cells. Loss of plasma membrane Ca2+-ATPase activity and decrease in superoxide production were not affected by glucose concentration, being seemingly determined by processes independent of both glycoxidation and energy depletion. These results point to the necessity of careful interpretation of data obtained in experiments, in which erythrocytes are subject to treatment with high glucose concentrations in vitro. PMID:26141922

  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. Oscillatory tank-treading motion of erythrocytes in shear flows

    PubMed Central

    Dodson, W. R.; Dimitrakopoulos, P.

    2013-01-01

    In this paper, we investigate the oscillatory dynamics of the tank-treading motion of healthy human erythrocytes in shear flows with capillary number Ca = O(1) and small to moderate viscosity ratios 0.01 ≤ λ ≤ 1.5. These conditions correspond to a wide range of surrounding medium viscosities (4 to 600 mPa s) and shear flow rates (2 to 560 s−1), and match those used in ektacytometry systems. For a given viscosity ratio, as the flow rate increases, the steady-state erythrocyte length L (in the shear plane) increases logarithmically while its depth W (normal to the shear plane) decreases logarithmically. In addition, the flow rate increase dampens the oscillatory erythrocyte inclination but not its lengths oscillations (which show relative variations of about 5–8%). For a given flow rate, as the viscosity ratio increases, the erythrocyte length L contracts while its depth W increases (i.e. the cell becomes less deformed) with small decrease in the lengths variations. The average orientation angle of the erythrocyte shows a significant decrease with the viscosity ratio as does the angle oscillation while the oscillation period increases. These trends continue in higher viscosity ratios resulting eventually in the transition from a (weakly oscillatory) tank-treading motion to a tumbling motion. Our computations show that the erythrocyte width S, which exists in the shear plane, is practically invariant in time, capillary number and viscosity ratio, and corresponds to a real cell thickness of about 2.5 μm. Comparison of our computational results with the predictions of (low degree-of-freedom) theoretical models and experimental findings, suggests that the energy dissipation due to the shape-memory effects is more significant than the energy dissipation due to the membrane viscosity. Our work shows that the oscillatory tank-treading motion can account for more than 50% of the variations found in ektacytometry systems; thus, researchers who wish to study inherent

  11. Oscillatory tank-treading motion of erythrocytes in shear flows.

    PubMed

    Dodson, W R; 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 ≤ λ ≤ 1.5. These conditions correspond to a wide range of surrounding medium viscosities (4-600 m Pa s) and shear flow rates (2-560 s(-1)), and match those used in ektacytometry systems. For a given viscosity ratio, as the flow rate increases, the steady-state erythrocyte length L (in the shear plane) increases logarithmically while its depth W (normal to the shear plane) decreases logarithmically. In addition, the flow rate increase dampens the oscillatory erythrocyte inclination but not its 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 2.5 μm. Comparison of our computational results with the predictions of (low degree-of-freedom) theoretical models and experimental findings, suggests that the energy dissipation due to the shape-memory effects is more significant than the energy dissipation due to the membrane viscosity. Our work shows that the oscillatory tank-treading motion can account for more than 50% of the variations found in ektacytometry systems; thus, researchers who wish to study inherent

  12. 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.

  13. Human erythrocytes are affected by the organochloride insecticide chlordane.

    PubMed

    Suwalsky, M; Rodríguez, C; Villena, F; Sotomayor, C P

    2005-05-01

    Chlordane is a widely used organochlorine insecticide. In order to evaluate its perturbing effect upon the morphology of human erythrocytes it was caused to interact with human red cells and molecular models of cell membranes. These consisted in bilayers of dimyristoylphosphatidylethanolamine (DMPE) and of dimyristoylphosphatidylcholine (DMPC), representative of phospholipid classes located in the inner and outer monolayers of the erythrocyte membrane, respectively. Scanning electron microscopy (SEM) observations indicated that this pesticide induced a significant alteration in the shape of the erythrocytes as they changed their discoid shape to spherocytes. According to the bilayer couple hypothesis, the shape changes induced in erythrocytes by foreign molecules are due to differential expansion of their two monolayers. The fact that chlordane produced spherocytes would indicate that the pesticide was equally located in the outer and the inner moieties of the red cell membrane. This conclusion was supported by the results obtained from X-ray diffraction studies. These showed that the hydrophobic and polar head regions of DMPC bilayers were perturbed when the insecticide was in a 1:10 molar ratio with respect to the lipid. These results were confirmed by the fluorescence experiments performed in DMPC large unilamellar vesicles (LUV). Chlordane produced a sharp decrease in the anisotropy and general polarization parameters in the 0-0.1 mM range, implying an increase in the fluidity at the acyl chain and polar region of DMPC. On the other hand, the bilayer structure of DMPE was perturbed in a fashion similar to that observed by X-ray diffraction in DMPC, a fact that explains the morphological change induced by chlordane to the human erythrocytes. PMID:15778003

  14. 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+}.

  15. [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.

  16. 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.

  17. 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.

  18. Uterine disorders in 50 pet rabbits.

    PubMed

    Künzel, Frank; Grinninger, Petra; Shibly, Sarina; Hassan, Jasmin; Tichy, Alexander; Berghold, Petra; Fuchs-Baumgartinger, Andrea

    2015-01-01

    Although the incidence of uterine disorders in pet rabbits is high there are only a few retrospective studies and case reports on genital tract disease in female rabbits. Uterine disorders were assessed in 50 pet rabbits. In 31 pet rabbits with suspected clinical uterine disease, medical records were further reviewed regarding clinical signs, diagnostic workup, treatment as well as the outcome itself. Uterine adenocarcinoma (54%) was most frequently diagnosed, followed by endometrial hyperplasia (26%). Serosanguineous vaginal discharge was the predominant clinical sign observed by the rabbit owners. In approximately 50% of the rabbits with suspected uterine disorders, abdominal palpation revealed enlarged and/or irregular masses in the caudoventral abdomen indicating uterine lesions. Out of 23 rabbits undergoing ovariohysterectomy, four were either euthanized or died shortly after surgery because they were clinically unstable. Overall, 80% of the ovariohysterectomized animals were still alive 6 mo after surgery. In female pet rabbits that are not breeding, either ovariohysterectomy should be performed at an early age or routine checks including ultrasonography of the abdomen are recommended on a regular basis.

  19. P. falciparum: merozoite surface protein-8 peptides bind specifically to human erythrocytes.

    PubMed

    Puentes, Alvaro; García, Javier; Ocampo, Marisol; Rodríguez, Luis; Vera, Ricardo; Curtidor, Hernando; López, Ramsés; Suarez, Jorge; Valbuena, John; Vanegas, Magnolia; Guzman, Fanny; Tovar, Diana; Patarroyo, Manuel E

    2003-07-01

    This work determined Plasmodium falciparum merozoite surface protein-8 (MSP-8) regions specifically binding to membrane surface receptors on human erythrocytes. Five high activity binding peptides (HABPs), whose binding to erythrocytes became saturable and sensitive on being treated with neuraminidase and chymotrypsin were identified from the MSP-8 protein. Those amino acids directly involved in interaction with erythrocytes were also determined for each one of the HABPs. Some of them specifically recognized 28, 46, and 73 kDa erythrocyte membrane proteins. Some HABPs inhibited in vitro P. falciparum merozoite invasion of erythrocytes by up to 98%, suggesting the MSP-8 protein's possible role in the invasion process.

  20. The physiologic role of erythrocytes in oxygen delivery and implications for blood storage.

    PubMed

    Benedik, Penelope S; Hamlin, Shannan K

    2014-09-01

    Erythrocytes are not just oxygen delivery devices but play an active metabolic role in modulating microvascular blood flow. Hemoglobin and red blood cell morphology change as local oxygen levels fall, eliciting the release of adenosine triphosphate and nitric oxide to initiate local vasodilation. Aged erythrocytes undergo physical and functional changes such that some of the red cell's most physiologically helpful attributes are diminished. This article reviews the functional anatomy and applied physiology of the erythrocyte and the microcirculation with an emphasis on how erythrocytes modulate microvascular function. The effects of cell storage on the metabolic functions of the erythrocyte are also briefly discussed.

  1. 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.

  2. 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.

  3. Phosphatidylserine externalization and procoagulant activation of erythrocytes induced by Pseudomonas aeruginosa virulence factor pyocyanin.

    PubMed

    Qadri, Syed M; Donkor, David A; Bhakta, Varsha; Eltringham-Smith, Louise J; Dwivedi, Dhruva J; Moore, Jane C; Pepler, Laura; Ivetic, Nikola; Nazi, Ishac; Fox-Robichaud, Alison E; Liaw, Patricia C; Sheffield, William P

    2016-04-01

    The opportunistic pathogen Pseudomonas aeruginosa causes a wide range of infections in multiple hosts by releasing an arsenal of virulence factors such as pyocyanin. Despite numerous reports on the pleiotropic cellular targets of pyocyanin toxicity in vivo, its impact on erythrocytes remains elusive. Erythrocytes undergo an apoptosis-like cell death called eryptosis which is characterized by cell shrinkage and phosphatidylserine (PS) externalization; this process confers a procoagulant phenotype on erythrocytes as well as fosters their phagocytosis and subsequent clearance from the circulation. Herein, we demonstrate that P. aeruginosa pyocyanin-elicited PS exposure and cell shrinkage in erythrocyte while preserving the membrane integrity. Mechanistically, exposure of erythrocytes to pyocyanin showed increased cytosolic Ca(2+) activity as well as Ca(2+) -dependent proteolytic processing of μ-calpain. Pyocyanin further up-regulated erythrocyte ceramide abundance and triggered the production of reactive oxygen species. Pyocyanin-induced increased PS externalization in erythrocytes translated into enhanced prothrombin activation and fibrin generation in plasma. As judged by carboxyfluorescein succinimidyl-ester labelling, pyocyanin-treated erythrocytes were cleared faster from the murine circulation as compared to untreated erythrocytes. Furthermore, erythrocytes incubated in plasma from patients with P. aeruginosa sepsis showed increased PS exposure as compared to erythrocytes incubated in plasma from healthy donors. In conclusion, the present study discloses the eryptosis-inducing effect of the virulence factor pyocyanin, thereby shedding light on a potentially important mechanism in the systemic complications of P. aeruginosa infection. PMID:26781477

  4. Abnormalities in the glycosphingolipid content of human Pk and p erythrocytes.

    PubMed Central

    Marcus, D M; Naiki, M; Kundu, S K

    1976-01-01

    Erythrocytes of the rare Pk phenotype lack the blood group P antigen, and p erythrocytes lack both P and Pk antigens. On the basis of immunological data we suggested previously that the P and Pk antigens are the glycosphingolipids globoside and trihexosyl ceramide, respectively, and we have now confirmed these designations by chemical analysis of erythrocytes lacking these antigens. The Pk erythrocytes contain only traces of globoside and have a marked excess of trihexosyl ceramide in comparison with normal erythrocytes. The p erythrocytes lack globoside and trihexosyl ceramide and contain an excess of lactosyl ceramide and other complex glycolipids. Our analyses of normal erythrocytes also revealed complex gangliosides with the approximate chromatographic mobilities of GD1b and GT1, and several gangliosides containing N-acetylglucosamine. Images PMID:1067617

  5. Serotyping scheme for Campylobacter jejuni and Campylobacter coli based on direct agglutination of heat-stable antigens.

    PubMed

    Frost, J A; Oza, A N; Thwaites, R T; Rowe, B

    1998-02-01

    Campylobacter is now the most frequently reported cause of gastrointestinal disease in England and Wales, yet few isolates are characterized beyond the genus level. The majority of isolates are Campylobacter jejuni (90%), with most of the remainder being Campylobacter coli. We describe an adaptation of the Penner serotyping scheme in which passive hemagglutination has been replaced by detection of heat-stable antigens by direct bacterial agglutination; absorbed antisera are used where appropriate. This scheme has been used to type 2,407 C. jejuni samples and 182 C. coli samples isolated in Wales between April 1996 and March 1997. Forty-seven C. jejuni serotypes were identified, with the 10 most prevalent serotypes accounting for 53% of the isolates tested; 19% of the isolates were untypeable. Only fifteen C. coli serotypes were identified, with three serotypes accounting for 69% of the isolates. This scheme provides a baseline for epidemiological studies of C. jejuni and C. coli.

  6. [Detection of toxoplasma-specific IgM antibodies--comparison with the ISAGA (immunosorbent agglutination assay) and immunofluorescence results].

    PubMed

    Saathoff, M; Seitz, H M

    1985-01-01

    Serum samples from 702 persons were examined for Toxoplasma-specific IgM-antibodies using the immunosorbent agglutination assay (ISAGA) and the immunofluorescence-test (IIFT). 250 samples showed a positive reaction in the Sabin-Feldman-test (SFT) with titers greater than or equal to 1: 1 024, 58% were positive in ISAGA and 36% in the IIFT. Samples of persons with acute Toxoplasma-infection, showed high titers in the ISAGA and SFT as well, even in cases where the IgM-IIFT was negative. SFT-negative sera and others with weakly-positive reactions and also rheuma-positive samples were negative in the ISAGA. It is discussed how far it is possible to determine the duration of the Toxoplasma infection by applying the ISAGA in combination with other antibody tests.

  7. [Standardization of neutralization tests using the COBL cell line and comparison with the particle agglutination test for measles serology].

    PubMed

    Korukluoğlu, Gülay; Yalçinkaya, Tülay; Ozkaya, Etem; Kurtoğlu, Demet; Gözalan, Ayşegül; Miyamura, Kikuko

    2002-04-01

    The aim of the present study was the detection and comparison of measles antibody titers with particle agglutination (PA) and neutralization (Nt) methods, in the sera samples of 364 subjects from different age groups. PA method was performed with a commercial test kit (Serodiameasles, Fujirebio Com. Japan), and Nt test which was standardized in this study, by using COBL (cord blood) cell lines, has been started to use in our laboratory as a reference method. As a result, antibody titers detected by PA were in parallel to the titers which detected by Nt test, and it was concluded that the differences in antibody titers would arise from the differences of test principles and viral antigens.

  8. The association between erythrocyte internal viscosity, protein non-enzymatic glycosylation and erythrocyte membrane dynamic properties in juvenile diabetes mellitus.

    PubMed Central

    Watala, C.; Witas, H.; Olszowska, L.; Piasecki, W.

    1992-01-01

    The association of intracellular viscosity of red blood cells and the dynamic properties of erythrocyte membranes in children suffering from diabetes has been investigated by means of ESR spectroscopy. It has been revealed that the slight decrease in the ratio hw/hs of maleimide bound to membrane protein-SH groups of erythrocytes in diabetes may ensue from the enhanced membrane protein immobilization in the plane of lipid bilayer. These alterations were accompanied by a corresponding increase in the relative rotational correlation time (tau c) of iodoacetamide spin label, thus suggesting that the conformational changes in membrane proteins may occur at both the intrinsic and more exposed thiol groups. The membranes of diabetic red blood cells were more glycosylated than those of relevant controls, and the extent of glycosylation was found to correlate significantly with h + 1/h0 and tau c (r = -0.652, P < 0.01 and r = 0.609, P < 0.01). Further, the conformational alterations in erythrocyte membranes from diabetic subjects were accompanied by a significant increase in the mobility parameter (h + 1/h0) of haemoglobin molecules in diabetic erythrocytes. The latter changes correlated well with the enhanced intracellular viscosity of diabetic red blood cells and the level of glycosylated haemoglobin. We conclude that the alterations in membrane lipid-protein interactions together with the increased glycosylation-derived internal viscosity may consequently imply altered viscoelastic properties of erythrocyte membranes and, underlying the impaired deformability of red blood cells in the diabetic state, contribute to the development of late diabetic sequelae. PMID:1329916

  9. Development and evaluation of a recombinant-glycoprotein-based latex agglutination test for rabies virus antibody assessment.

    PubMed

    Jemima, Ebenezer Angel; Manoharan, Seeralan; Kumanan, Kathaperumal

    2014-08-01

    The measurement of neutralizing antibodies induced by the glycoprotein of rabies virus is indispensable for assessing the level of neutralizing antibodies in animals or humans. A rapid fluorescent focus inhibition test (RFFIT) has been approved by WHO and is the most widely used method to measure the virus-neutralizing antibody content in serum, but a rapid test system would be of great value to screen large numbers of serum samples. To develop and evaluate a latex agglutination test (LAT) for measuring rabies virus antibodies, a recombinant glycoprotein was expressed in an insect cell system and purified, and the protein was coated onto latex beads at concentrations of 0.1, 0.25, 0.5, 0.75, and 1 mg/ml to find out the optimal concentration for coating latex beads. It was found that 0.5 mg/ml of recombinant protein was optimal for coating latex beads, and this concentration was used to sensitize the latex beads for screening of dog serum samples. Grading of LAT results was done with standard reference serum with known antibody titers. A total of 228 serum samples were tested, out of which 145 samples were positive by both RFFIT and LAT, and the specificity was found to be 100 %. In RFFIT, 151 samples were positive, the sensitivity was found to be 96.03 %, and the accuracy/concordance was found to be 97.39 %. A rapid field test-a latex agglutination test (LAT)-was developed and evaluated for rabies virus antibody assessment using recombinant glycoprotein of rabies virus expressed in an insect cell system.

  10. Immunoglobulin-Mediated Agglutination of and Biofilm Formation by Escherichia coli K-12 Require the Type 1 Pilus Fiber

    PubMed Central

    Orndorff, Paul E.; Devapali, Aditya; Palestrant, Sarah; Wyse, Aaron; Everett, Mary Lou; Bollinger, R. Randal; Parker, William

    2004-01-01

    The binding of human secretory immunoglobulin A (SIgA), the primary immunoglobulin in the gut, to Escherichia coli is thought to be dependent on type 1 pili. Type 1 pili are filamentous bacterial surface attachment organelles comprised principally of a single protein, the product of the fimA gene. A minor component of the pilus fiber (the product of the fimH gene, termed the adhesin) mediates attachment to a variety of host cell molecules in a mannose inhibitable interaction that has been extensively described. We found that the aggregation of E. coli K-12 by human secretory IgA (SIgA) was dependent on the presence of the pilus fiber, even in the absence of the mannose specific adhesin or in the presence of 25 mM α-CH3Man. The presence of pilus without adhesin also facilitated SIgA-mediated biofilm formation on polystyrene, although biofilm formation was stronger in the presence of the adhesin. IgM also mediated aggregation and biofilm formation in a manner dependent on pili with or without adhesin. These findings indicate that the pilus fiber, even in the absence of the adhesin, may play a role in biologically important processes. Under conditions in which E. coli was agglutinated by SIgA, the binding of SIgA to E. coli was not increased by the presence of the pili, with or without adhesin. This observation suggests that the pili, with or without adhesin, affect factors such as cell surface rigidity or electrostatic repulsion, which can affect agglutination but which do not necessarily determine the level of bound immunoglobulin. PMID:15039312

  11. 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.

  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, 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...

  14. 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...

  15. 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...

  16. Clinical aspects of lagomorph dental anatomy: the rabbit (oryctolagus cuniculus).

    PubMed

    Crossley, D A

    1995-12-01

    The lagomorphs most commonly encountered as pets are rabbits. There are many breeds of domestic rabbit, varying from dwarf varieties with an adult weight of under one kilogram to giants weighing 10 kg. This article provides a working knowledge of the dental anatomy and physiology of rabbits so that veterinarians can interpret clinical and radiographic findings when investigating rabbits with suspected dental disease.

  17. 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...

  18. 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

  19. Is Peroxiredoxin II's peroxidase activity strongly inhibited in human erythrocytes?

    PubMed

    Benfeitas, Rui; Selvaggio, Gianluca; Antunes, Fernando; Coelho, Pedro; Salvador, Armindo

    2014-10-01

    H2O2 elimination in human erythrocytes is mainly carried out by catalase (Cat), glutathione peroxidase (GPx1) and the more recently discovered peroxiredoxin 2 (Prx2). However, the contribution of Prx2 to H2O2 consumption is still unclear. Prx2's high reactivity with H2O2 (kPrx2=10×10(7) M(-1)s(-1), kCat =7×10(7) M(-1)s(-1), kGPx1 =4×10(7) M(-1)s(-1)) and high abundance ([Prx2]= 570µM, [Cat]= 32µM, [GPx1]= 1µM) suggest that under low H2O2 supply rates it should consume >99% of the H2O2. However, extensive evidence indicates that in intact erythrocytes Prx2 contributes no more than Cat to H2O2 consumption. In order for this to be attained, Prx2's effective rate constant with H2O2would have to be just ~10(5) M(-1)s(-1), much lower than that determined in multiple experiments with the purified proteins. Nevertheless, nearly all Prx2 is oxidized within 1min of exposing erythrocytes to a H2O2 bolus, which is inconsistent with an irreversible inhibition. A mathematical model of the H2O2 metabolism in human erythrocytes [Benfeitas et al. (2014) Free Radic. Biol. Med.] where Prx2 either has a low kPrx2 or is subject to a strong (>99%) but readily reversible inhibition achieves quantitative agreement with detailed experimental observations of the responses of the redox status of Prx2 in human erythrocytes and suggests functional advantages of this design (see companion abstract). By contrast, a variant where Prx2 is fully active with kPrx2=10(8) M(-1)s(-1) shows important qualitative discrepancies. Altogether, these results suggest that Prx2's peroxidase activity is strongly inhibited in human erythrocytes. We acknowledge fellowship SFRH/BD/51199/2010, grants PEst-C/SAU/LA0001/2013-2014, PEst-OE/QUI/UI0612/2013, PEst-OE/QUI/UI0313/2014, and FCOMP-01-0124-FEDER-020978 (PTDC/QUI-BIQ/119657/2010) co-financed by FEDER through the COMPETE program and by FCT.

  20. Partial purification and characterization of an actin-bundling protein, band 4.9, from human erythrocytes.

    PubMed

    Siegel, D L; Branton, D

    1985-03-01

    Band 4.9 (a 48,000-mol-wt polypeptide) has been partially purified from human erythrocyte membranes. In solution, band 4.9 polypeptides exist as trimers with an apparent molecular weight of 145,000 and a Stokes radius of 50 A. Electron microscopy shows that the protein is a three-lobed structure with a radius slightly greater than 50 A. When gel-filtered rabbit muscle actin is polymerized in the presence of band 4.9, actin bundles are generated that are similar in appearance to those induced by "vinculin" or fimbrin. The bundles appear brittle and when they are centrifuged small pieces of filaments break off and remain in the supernatant. At low band 4.9 to actin molar ratios (1:30), band 4.9 lowers the apparent steady-state low-shear falling ball viscosity by sequestering filaments into thin bundles; at higher ratios, the bundles become thicker and obstruct the ball's movement leading to an apparent increase in steady-state viscosity. Band 4.9 increases the length of the lag phase and decreases the rate of elongation during actin polymerization as measured by high-shear Ostwald viscometry or by the increase in the fluorescence of pyrene-labeled actin. Band 4.9 does not alter the critical actin monomer concentration. We hypothesize that band 4.9, together with actin, erythrocyte tropomyosin, and spectrin, forms structures in erythroid precursor cells analogous to those formed by fimbrin, actin, tropomyosin, and TW 260/240 in epithelial brush borders. During erythroid development and enucleation, the actin filaments may depolymerize up to the membrane, leaving a membrane skeleton with short stubs of actin bundled by band 4.9 and cross-linked by spectrin. PMID:3882722

  1. Strain differentiation of Neisseria gonorrhoeae by reverse passive hemagglutination.

    PubMed Central

    Armstrong, A S; Mathias, J R; DeYoung, M I; Hirata, A A

    1979-01-01

    A reverse passive hemagglutination test that utilizes human erythrocytes coated with antibody to gonococci was developed to distinguish differences among 11 strains of Neisseria gonorrhoeae. Different rabbits were immunized with each strain of gonococcus. Antibody was purified by passing antiserum over an immunoadsorbent column containing homologous cell walls trapped in a cross-linked polyacrylamide gel. Antibody, after absorption with N. meningitidis, was used for coating 11 individual suspensions of erythrocytes, each with antibody to one gonococcal strain. The panel of coated erythrocytes was added to microtiter trays containing dilutions of homologous bacterial lysate and lysates from 10 heterologous strains. Agglutination titers were highest with homologous lysates, although cross-reactions occurred among some heterologous lysates. Lysates of nongonococcal Neisseria species and of other genera did not agglutinate coated erythrocytes. The reverse passive hemagglutination test can be a useful procedure to distinguish differences among strains of N. gonorrhoeae. Images PMID:110695

  2. Properties of Streptococcus mutans Grown in a Synthetic Medium: Binding of Glucosyltransferase and In Vitro Adherence, and Binding of Dextran/Glucan and Glycoprotein and Agglutination

    PubMed Central

    Wu-Yuan, Christine D.; Tai, Stella; Slade, Hutton D.

    1979-01-01

    The influence of culture media on various properties of Streptococcus mutans was investigated. Strains of S. mutans (serotypes c, d, f, and g) were grown in a complex medium (Todd-Hewitt broth [THB]) or a synthetic medium (SYN). The SYN cells, in contrast to THB cells, did not bind extracellular glucosyltransferase and did not produce in vitro adherence. Both types of cells possessed constitutive levels of glucosyltransferase. B13 cells grown in SYN plus invertase-treated glucose possessed the same level of constitutive enzyme as THB cells. In contrast to THB cells, the SYN cells of seven serotype strains did not agglutinate upon the addition of high-molecular-weight dextran/glucan. Significant quantities of lower-molecular-weight (2 × 104 or 7 × 104) dextran and B13 glucan were bound by SYN cells. SYN cells agglutinated weakly in anti-glucan serum (titers, 0 to 16), whereas THB cells possessed titers of 32 to 256. Evidence for the existence of a second binding site in agglutination which does not possess a glucan-like polymer has been obtained. B13 cells grown in invertase-treated THB agglutinated to the same degree as normal THB cells. The nature of this site is unknown. SYN cells possess the type-specific polysaccharide antigen. B13 cells did not bind from THB a glycoprotein which reacts with antisera to the A, B, or T blood group antigens or which allows agglutination upon the addition of dextran. The results demonstrate that S. mutans grown in a chemically defined medium possesse markedly different biochemical and biological activities than cells grown in a complex organic medium. PMID:457252

  3. Heat transport in laminar flow of erythrocyte suspensions.

    PubMed

    Ahuja, A S

    1975-07-01

    Measurements of thermal conductivity were made in laminar flow of dog and turkey erythrocyte suspensions in a stainless stell tube of about 1 mm ID. These measurements were independent of the shear rate, showing that the red cell motion relative to plasma in flowing blood had no effect on the heat transfer. Measurements of thermal conductivity were further made in suspensions of polystyrene spheres of 100 mum and were found to be dependent upon the shear rate. The Graetz solution corresponding to uniform wall temperature was used for determining the value of thermal conductivity in an apparatus calibrated with tap water. The overall accuracy of the results is within 10%. A model based on the particle rotation with the entrained fluid is proposed. It is pointed out that the diffusion of platelets, red cells, and possibly plasma proteins (such as fibrinogen) will be augmented if they happen to be in the hydrodynamic field of rotating erythrocytes. PMID:1150598

  4. Effects of ascorbate fatty ester derivatives on erythrocyte membrane lipoperoxidation.

    PubMed

    Spengler, M I; Rasia, M; Palma, S; Allemandi, D

    2011-01-01

    6-O-alkyl ascorbic acid esters (ASC(n)) are amphiphilic molecules that behave as surfactants in aqueous solution. ASC(n) have shown some physical and rheological properties that suggest a potential utility as drug carriers. The present paper aims to evaluate the effect of ASC(n) on erythrocyte properties in order to get information regarding the relationship between osmotic fragility, erythrocyte deformability and membrane lipoperoxidation process. The assays were performed at the following concentrations: the critical micelar concentration (CMC), producing 10% hemolysis (CH(10)) and producing 50% hemolysis (CH(50)). We observed that ASC(n) (ASC(8), ASC(10) and ASC(12)), at concentration nearby CMC, neither affected cell deformability nor produced lipoperoxidation. Nevertheless, at higher concentrations (CH(10) and CH(50)), the RBCs incubated with ASC(n) were affected by a significant and progressive loss of deformability, simultaneously with an increase of osmotic fragility and membrane lipoperoxidation.

  5. Human erythrocytes as nanoparticle carriers for magnetic particle imaging.

    PubMed

    Markov, D E; Boeve, H; Gleich, B; Borgert, J; Antonelli, A; Sfara, C; Magnani, M

    2010-11-01

    The potential of red blood cells (RBCs) loaded with iron oxide nanoparticles as a tracer material for magnetic particle imaging (MPI) has been investigated. MPI is an emerging, quantitative medical imaging modality which holds promise in terms of sensitivity in combination with spatial and temporal resolution. Steady-state and dynamic magnetization measurements, supported by semi-empirical modeling, were employed to analyze the MPI signal generation using RBCs as novel biomimetic constructs. Since the superparamagnetic iron oxide (SPIO) bulk material that is used in this study contains nanoparticles with different sizes, it is suggested that during the RBC loading procedure, a preferential entrapment of nanoparticles with hydrodynamic diameter ≤60 nm occurs by size-selection through the erythrocyte membrane pores. This affects the MPI signal of an erythrocyte-based tracer, compared to bulk. The reduced signal is counterbalanced by a higher in vivo stability of the SPIO-loaded RBCs constructs for MPI applications.

  6. Cell Electrofusion in Centrifuged Erythrocyte Pellets Assessed by Dielectric Spectroscopy.

    PubMed

    Asami, Koji

    2016-04-01

    We have characterized cell electrofusion in cell pellets by dielectric spectroscopy. Cell pellets were formed from horse erythrocyte suspensions by centrifugation and were subjected to intense AC pulses. The dielectric spectra of the pellets were measured over a frequency range of 10 Hz to 10 MHz. The application of AC pulses caused low-frequency (LF) dielectric relaxation below about 100 kHz. The LF dielectric relaxation was markedly affected not only by pretreatment of cells at 50 °C, which disrupts the spectrin network of erythrocytes, but also by the parameters of the AC pulses (frequency of the sine wave and repeat count of the pulses). The occurrence of the LF dielectric relaxation was qualitatively accounted for by modeling fusion products in the pellet by prolate spheroidal cells whose long axes run parallel to the applied electric field.

  7. 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

  8. Heat transport in laminar flow of erythrocyte suspensions.

    PubMed

    Ahuja, A S

    1975-07-01

    Measurements of thermal conductivity were made in laminar flow of dog and turkey erythrocyte suspensions in a stainless stell tube of about 1 mm ID. These measurements were independent of the shear rate, showing that the red cell motion relative to plasma in flowing blood had no effect on the heat transfer. Measurements of thermal conductivity were further made in suspensions of polystyrene spheres of 100 mum and were found to be dependent upon the shear rate. The Graetz solution corresponding to uniform wall temperature was used for determining the value of thermal conductivity in an apparatus calibrated with tap water. The overall accuracy of the results is within 10%. A model based on the particle rotation with the entrained fluid is proposed. It is pointed out that the diffusion of platelets, red cells, and possibly plasma proteins (such as fibrinogen) will be augmented if they happen to be in the hydrodynamic field of rotating erythrocytes.

  9. Human erythrocytes as nanoparticle carriers for magnetic particle imaging.

    PubMed

    Markov, D E; Boeve, H; Gleich, B; Borgert, J; Antonelli, A; Sfara, C; Magnani, M

    2010-11-01

    The potential of red blood cells (RBCs) loaded with iron oxide nanoparticles as a tracer material for magnetic particle imaging (MPI) has been investigated. MPI is an emerging, quantitative medical imaging modality which holds promise in terms of sensitivity in combination with spatial and temporal resolution. Steady-state and dynamic magnetization measurements, supported by semi-empirical modeling, were employed to analyze the MPI signal generation using RBCs as novel biomimetic constructs. Since the superparamagnetic iron oxide (SPIO) bulk material that is used in this study contains nanoparticles with different sizes, it is suggested that during the RBC loading procedure, a preferential entrapment of nanoparticles with hydrodynamic diameter ≤60 nm occurs by size-selection through the erythrocyte membrane pores. This affects the MPI signal of an erythrocyte-based tracer, compared to bulk. The reduced signal is counterbalanced by a higher in vivo stability of the SPIO-loaded RBCs constructs for MPI applications. PMID:20959685

  10. Recent advancements in erythrocytes, platelets, and albumin as delivery systems.

    PubMed

    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.

  11. 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.

  12. 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.

  13. 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.

  14. Structural specificity of serotonin effect on human erythrocyte fragility.

    PubMed

    Gilboa-Garber, N; Kirstein-Segal, R

    1998-08-01

    Serotonin, a neurotransmitter and vasoconstrictor, affects various cell properties. We have analyzed the importance of its structural components for its extensive effect on human erythrocyte fragility, using its O- and N-linked derivatives and related compounds. The results presented in this communication indicate that the amino group, free of adjacent negative charges, and the hydroxyl group are indispensable for the serotonin-induced increase in red blood cell fragility. PMID:9758719

  15. Structural alterations of erythrocyte membrane components induced by exhaustive exercise.

    PubMed

    Brzeszczynska, Joanna; Pieniazek, Anna; Gwozdzinski, Lukasz; Gwozdzinski, Krzysztof; Jegier, Anna

    2008-12-01

    Physical exercise was used as a model of the physiological modulator of free radical production to examine the effects of exercise-induced oxidative modifications on the physico-biochemical properties of erythrocyte membrane. The aim of our work was to investigate conformational changes of erythrocyte membrane proteins, membrane fluidity, and membrane susceptibility to disintegration. Venous blood was taken before, immediately after, and 1 h after an exhaustive incremental cycling test (30 W.min-1 ramp), performed by 11 healthy untrained males on balanced diets (mean age, 22 +/- 2 years; mean body mass index, 25 +/- 4.5 kg.m-2). In response to this exercise, individual maximum heart rate was 195 +/- 12 beats.min-1 and maximum wattage was 292 +/- 27 W. Electron paramagnetic resonance spectroscopy was used to investigate alterations in membrane proteins and membrane dynamics, and to measure production of radical species. The reducing potential of plasma (RPP) was measured using the reduction of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the ferric-reducing ability of plasma. Exercise induced decreases in erythrocyte membrane fluidity in the polar region (p < 0.0001) and alterations in the conformational state of membrane proteins (p < 0.05). An increase in RPP was observed immediately after exercise (p < 0.001), with a further increase 1 h postexercise (p < 0.0001). Supporting measurements of lipid peroxidation showed an increase in thiobarbituric acid reactive substances immediately after exercise (p < 0.05) and at 1 h of recovery (p < 0.001); however, free radicals were not detected. Results indicate the existence of early postexercise mild oxidative stress after single-exercise performance, which induced structural changes in erythrocyte membrane components (protein aggregation) and in the membrane organization (lipids rigidization) that followed lipid peroxidation but did not lead to cellular hemolysis.

  16. Sodium and potassium transport in trout (Salmo gairdneri) erythrocytes.

    PubMed Central

    Bourne, P K; Cossins, A R

    1984-01-01

    The principal pathways of Na+ and K+ transport in trout erythrocytes have been characterized. Approximately 50% of K+ influx in steady-state erythrocytes was inhibited by ouabain (1 mM) and 46% by furosemide (1 mM). Furosemide-sensitive K+ influx was a saturable function of external K+ concentration with a Km of 25 mM. This flux component was also inhibited by SITS (4-acetamido-4'-isothiocyanatostilbene-2'2-disulphonate) (concentration required for 50% inhibition, I50 = 7.6 X 10(-6)M) and by the removal of external Cl-. An increase in cell volume stimulated furosemide-sensitive K+ influx and cell shrinkage inhibited this flux. K+ efflux was mainly furosemide-sensitive (64% of total). This pathway was unaffected by variations in extracellular K+ concentration and is therefore not exchange diffusion. However, it was affected by variations in cell volume in a similar way to the furosemide-sensitive K+ influx. Na+ influx was only slightly sensitive to furosemide (13% of total) but this component was very sensitive to changes in cell volume; decreased cell volume increased Na+ influx whilst increased cell volume inhibited Na+ influx. Furosemide-sensitive K+ influx was unaffected by variations in external Na+ concentration. Similarly, furosemide-sensitive Na+ influx was unaffected by variations in external K+ concentration. This indicates that the passive influxes of Na+ and K+ were not coupled, in contrast to the situation in avian erythrocytes. The opposite effects of cell volume upon passive Na+ and K+ fluxes are in good agreement with the net movements of these cations during volume regulation in erythrocytes of the flounder (Cala, 1977) and the toadfish (Lauf, 1982). PMID:6707960

  17. Are polyphosphoinositides associated with glycophorin in human erythrocyte membranes?

    PubMed

    Shukla, S D; Coleman, R; Finean, J B; Michell, R H

    1979-05-01

    Glycophorin prepared by a lithium di-iodosalicylate-extraction/phenol-partition method was rich in polyphosphoinositides (phosphatidyl-myo-inositol 4-phosphate and phosphatidyl-myo-inositol 4,5-bisphosphate), but glycophorin extracted by Triton X-100 showed no such enrichment. The enrichment observed in the former preparations appeared not to be caused by pre-existing association between glycophorin and polyphosphoinositides in the human erythrocyte membrane, but to be largely a consequence of the preparative procedures.

  18. Isolation and some properties of glycated D-glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle.

    PubMed Central

    He, R Q; Yang, M D; Zheng, X; Zhou, J X

    1995-01-01

    Glycated D-glyceraldehyde-3-phosphate dehydrogenases (GAPDH) from rabbit muscle and human erythrocytes have been investigated. The specific activity of the non-glycated GAPDH from rabbit muscle is approx. 180 units. (One unit is defined as the specific activity required to convert 1 microM of substrate/min per mg of enzyme.) The activity of the glycated enzyme, consisting of two sugars per tetramer, is lower than that of the non-glycated GAPDH. Non-enzymic transamination of the N-termini of glycated GAPDH (gGAPDH) indicates that they are not blocked by glycation. The rate of modification of thiols (Cys-149) with 5,5'-dithiobis-(2-nitrobenzoic acid) was greater for the glycated than the non-glycated enzymes. The rate of modification of amino groups of Lys residues of gGAPDH with o-phthalaldehyde was greater for the non-glycated enzyme. In 0.18 M guanidine-HC1 solution, the emission intensity at 410 nm of a fluorescent NAD+ derivative introduced into the active site decreased to 80%, whereas that of gGAPDH decreased to 50%. This suggests that the glycated sites are near the active site; glycation of the enzyme leads to a change of the microenvironment of Cys-149, alters the conformation of the active site and decreases the activity. Images Figure 1 PMID:7619048

  19. 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.

  20. 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.

  1. 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. PMID:26929599

  2. 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

  3. Erythrocyte deformability - A partner of the inflammatory response.

    PubMed

    Silva-Herdade, Ana Santos; Andolina, Giulia; Faggio, Caterina; Calado, Ângelo; Saldanha, Carlota

    2016-09-01

    We aim to establish an in vivo animal model of acute inflammation using PAF (platelet activating factor) as inflammatory agent and to study the erythrocyte deformability changes induced by the inflammatory response. Counting the number of rolling and adherent neutrophils to endothelium after 2, 4 and 6h of intrascrotal injection of PAF we showed the induction of an inflammatory state. Blood samples are collected in order to measure the erythrocyte deformability and to quantify NO efflux from the red blood cells (RBCs). The results show an increased number of rolling and adherent neutrophils after 2h and 4h of inflammation as well as decreased values of erythrocyte deformability in the same time-points. This result is in line with the need of a low blood viscosity to the recruitment process that will improve leukocyte migration towards the endothelial wall. NO efflux from RBCs is also affected by the inflammatory response at the first hours of inflammation. This animal model demonstrates in vivo the association between an acute inflammatory response and the rheological properties of the blood, namely the RBCs deformability. For those reasons we consider this as an adequate model to study acute inflammatory responses as well as hemorheological parameters. PMID:27142964

  4. 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.

  5. 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.

  6. Modulation Effects of Curcumin on Erythrocyte Ion-Transporter Activity

    PubMed Central

    Singh, Prabhakar; Rizvi, Syed Ibrahim

    2015-01-01

    Curcumin ((1E,6E)-1,7-Bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione), the yellow biphenolic pigment isolated from turmeric (Curcuma longa), has various medicinal benefits through antioxidation, anti-inflammation, cardiovascular protection, immunomodulation, enhancing of the apoptotic process, and antiangiogenic property. We explored the effects of curcumin in vitro (10−5 M to 10−8 M) and in vivo (340 and 170 mg/kg b.w., oral) on Na+/K+ ATPase (NKA), Na+/H+ exchanger (NHE) activity, and membrane lipid hydroperoxides (ROOH) in control and experimental oxidative stress erythrocytes of Wistar rats. As a result, we found that curcumin potently modulated the membrane transporters activity with protecting membrane lipids against hydro-peroxidation in control as well as oxidatively challenged erythrocytes evidenced by stimulation of NKA, downregulation of NHE, and reduction of ROOH in the membrane. The observed results corroborate membrane transporters activity with susceptibility of erythrocyte membrane towards oxidative damage. Results explain the protective mechanism of curcumin against oxidative stress mediated impairment in ions-transporters activity and health beneficial effects. PMID:26421014

  7. 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.

  8. Electro-orientation of ellipsoidal erythrocytes. Theory and experiment.

    PubMed Central

    Miller, R D; Jones, T B

    1993-01-01

    The frequency-dependent orientation of human and llama erythrocytes suspended in isotonic solutions and subjected to linearly polarized electric fields is examined. Human erythrocytes may be represented as oblate spheroids (3.9:3.9:1.1 microns) with two distinguishable orientations, while the llama cells are approximated as ellipsoids with three distinct axes (4.0:2.0:1.1 microns). Under appropriate experimental conditions, both orientations of the human cells and all three orientations of the llama cells are observed. A theoretical cell model which accounts for the membrane as a thin confocal layer of ideal capacitance is used to predict the orientational spectra. The predicted spectra compare favorably in frequency range and orientational sequence with experimental data. Estimates for cell internal conductivity and permittivity are obtained by adjusting the values of these important parameters to achieve the closet fit of the theoretical curves to the data. By the use of this method, the internal conductivity of llama erythrocytes is estimated to be 0.26 S/m (+/- 20%), while the effective internal dielectric constant and conductivity of Euglena gracilis are estimated to be 120 (+/- 10%) and 0.43 S/m (+/- 20%), respectively. PMID:8324193

  9. Diet of racing sled dogs affects erythrocyte depression by stress.

    PubMed

    Adkins, T O; Kronfeld, D S

    1982-09-01

    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

  10. 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. PMID:27071054

  11. 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.

  12. EXPERIMENTAL SYPHILIS IN THE RABBIT

    PubMed Central

    Brown, Wade H.; Pearce, Louise

    1920-01-01

    From a study of the phenomena of the primary infection on the one hand, and the phenomena of local spread, or dissemination, on the other, it is seen that a multiplicity of lesions develops in the testicle and scrotum of the rabbit which have much the same characteristics irrespective of their origin. Some of these lesions are clearly recognizable as primary lesions or parts of a primary reaction to infection, while others are just as clearly the results of dissemination of the virus from a primary focus of infection or correspond with lesions which are commonly spoken of as secondary lesions. The effort to draw a sharp line of distinction between these two groups of lesions or between a primary and a secondary stage of infection in the rabbit, however, would be largely an arbitrary procedure. The fact is that the tissues of the scrotum and testicle of the rabbit constitute favorable surroundings for the localization and development of pallidum infections. Under ordinary circumstances, a large part of the reaction to infection which expresses itself in the formation of lesions recognizable by ordinary methods of examination takes place in these tissues. These lesions present certain broad and general characteristics without regard to whether they are primary or secondary in origin; the reaction is merely a reaction to a syphilitic infection which in either case may assume the most diverse character. Further, it would appear that in rabbits infected with such strains of Treponema pallidum as we have used, the virus is never confined to the area occupied by the so called primary lesion, or chancre, but always spreads and always gives rise to a regional adenopathy. There may be no lesions to indicate the progress of this dissemination, but an examination of the inguinal nodes shows that dissemination occurs very soon after inoculation, and a pallidum reaction may be detected in these glands even before infection can be recognized in the scrotum. Subsequently lesions

  13. Classical conditioning of the nictitating membrane response of the rabbit. III. Connections of cerebellar lobule HVI.

    PubMed

    Yeo, C H; Hardiman, M J; Glickstein, M

    1985-01-01

    We report the connections of cerebellar cortical lobule HVI in the rabbit. We have studied the anterograde and retrograde transport of wheatgerm-agglutinated horseradish peroxidase (WGA-HRP) following its injection into HVI to reveal efferent and afferent connections. All of the cases showed strong anterograde transport to the anterior interpositus nucleus (AIP) - indicating that this is the major efferent target of HVI. Retrogradely labelled cells were found in the inferior olivary, spinal trigeminal, lateral reticular, inferior vestibular and pontine nuclei. Within the olive, the medial part of the rostral dorsal accessory olive (DAO) and the adjacent medial part of the principal olive (PO) were consistently labelled in all cases. This area is known to receive somatosensory information from the face and neck. There was no projection to the hemispheral part of lobule VI from visual parts of the olive within the dorsal cap and medial parts of the medial accessory olive. Likely sources of visual and auditory information to HVI are the dorsolateral basilar pontine nuclei and nucleus reticularis tegmenti pontis, which were densely labelled in all cases. These anatomical findings are consistent with the suggestion that, during NMR conditioning, information related to the periorbital shock unconditional stimulus (US) may be provided by climbing fibres to HVI and light and white noise conditional stimulus (CS) information may be supplied by pontine mossy fibres. PMID:4043270

  14. Electrical constants of arterially perfused rabbit papillary muscle.

    PubMed Central

    Kléber, A G; Riegger, C B

    1987-01-01

    1. Right ventricular rabbit papillary muscles were arterially perfused with a mixture of Tyrode solution, bovine erythrocytes, dextran and albumin. In the recording chamber, they were surrounded by a H2O-saturated atmosphere of O2 and CO2 which served as an electrical insulator. 2. Conduction velocity and passive electrical properties were determined from intra- and extracellular potentials measured during excitation and during flow of subthreshold current. 3. The propagation of the action potential was linear along the muscle at a velocity of 55.6 cm/s. The extracellular wave-front voltage was 51.5 mV. 4. The following values for passive cable properties were obtained: (i) a ratio of extra- to intracellular longitudinal resistance of 1.2; (ii) an extracellular specific resistance (Ro) of 63 omega cm; (iii) an intracellular specific resistance (Ri) of 166 omega cm; (iv) a space constant lambda of 0.357 mm; (v) a membrane time constant tau of 2.57 ms. The space constant lambda* recalculated for zero extracellular resistance was 0.528 mm. 5. Arresting perfusion with drop of perfusion pressure was associated with an immediate increase of the extracellular longitudinal resistance by 35% and a decrease of conduction velocity by 13%. 6. The present results demonstrate the important contribution of the extracellular resistance to electrotonic interaction and propagation in densely packed myocardial tissue. Moreover, changes in perfusion pressure are associated with changes in extracellular resistance, probably as a consequence of changes in intravascular volume. PMID:3656162

  15. Rabbit Models for Studying Human Infectious Diseases

    PubMed Central

    Peng, Xuwen; Knouse, John A; Hernon, Krista M

    2015-01-01

    Using an appropriate animal model is crucial for mimicking human disease conditions, and various facets including genetics, anatomy, and pathophysiology should be considered before selecting a model. Rabbits (Oryctolagus cuniculus) are well known for their wide use in production of antibodies, eye research, atherosclerosis and other cardiovascular diseases. However, a systematic description of the rabbit as primary experimental models for the study of various human infectious diseases is unavailable. This review focuses on the human infectious diseases for which rabbits are considered a classic or highly appropriate model, including AIDS (caused by HIV1), adult T-cell leukemia–lymphoma (human T-lymphotropic virus type 1), papilloma or carcinoma (human papillomavirus) , herpetic stromal keratitis (herpes simplex virus type 1), tuberculosis (Mycobacterium tuberculosis), and syphilis (Treponema pallidum). In addition, particular aspects of the husbandry and care of rabbits used in studies of human infectious diseases are described. PMID:26678367

  16. Eyeblink conditioning in the developing rabbit

    PubMed Central

    Brown, Kevin L.; Woodruff-Pak, Diana S.

    2011-01-01

    Eyeblink classical conditioning in pre-weanling rabbits was examined in the present study. Using a custom lightweight headpiece and restrainer, New Zealand white littermates were trained once daily in 400 ms delay eyeblink classical conditioning from postnatal days (PD) 17–21 or PD 24–28. These ages were chosen because eyeblink conditioning emerges gradually over PD 17–24 in rats (Stanton, Freeman, & Skelton, 1992), another altricial species with neurodevelopmental features similar to those of rabbits. Consistent with well-established findings in rats, rabbits trained from PD 24–28 showed greater conditioning relative to littermates trained from PD 17–21. Both age groups displayed poor retention of eyeblink conditioning at retraining one month after acquisition. These findings are the first to demonstrate eyeblink conditioning in the developing rabbit. With further characterization of optimal conditioning parameters, this preparation may have applications to neurodevelopmental disease models as well as research exploring the ontogeny of memory. PMID:21953433

  17. Rabbit Models for Studying Human Infectious Diseases.

    PubMed

    Peng, Xuwen; Knouse, John A; Hernon, Krista M

    2015-12-01

    Using an appropriate animal model is crucial for mimicking human disease conditions, and various facets including genetics, anatomy, and pathophysiology should be considered before selecting a model. Rabbits (Oryctolagus cuniculus) are well known for their wide use in production of antibodies, eye research, atherosclerosis and other cardiovascular diseases. However, a systematic description of the rabbit as primary experimental models for the study of various human infectious diseases is unavailable. This review focuses on the human infectious diseases for which rabbits are considered a classic or highly appropriate model, including AIDS (caused by HIV1), adult T-cell leukemia-lymphoma (human T-lymphotropic virus type 1), papilloma or carcinoma (human papillomavirus) , herpetic stromal keratitis (herpes simplex virus type 1), tuberculosis (Mycobacterium tuberculosis), and syphilis (Treponema pallidum). In addition, particular aspects of the husbandry and care of rabbits used in studies of human infectious diseases are described. PMID:26678367

  18. Supplementation of the black rice outer layer fraction to rabbits decreases atherosclerotic plaque formation and increases antioxidant status.

    PubMed

    Ling, Wen Hua; Wang, Lin Llin; Ma, Jing

    2002-01-01

    The influence of the supplementation of black and white rice outer layer fractions on atherosclerotic plaque formation induced by hypercholesterolemia was investigated in rabbits. Male rabbits (n = 32) were randomly divided into four groups. They were fed nonpurified diet (normal group), a lard (3.5 g/100 g) with high cholesterol (0.5 g/100 g) diet (HC group); the HC diet with 5 g/100 g white rice outer layer fraction (WRF group); or the HC diet with 5 g/100 g black rice outer layer fraction (BRF) for 2 mo. Blood samples were collected for determination of lipid concentration and oxidative and antioxidative status variables, and aortae were taken for the assessment of atherosclerotic plaques. The atherosclerotic plaque area in rabbits fed the BRF diet was 66% lower than that of the HC or WRF rabbits (P < 0.001). Supplementation of the black rice outer layer significantly (P < 0.05) lowered aortic 8-hydroxy-2'-deoxyguanosine (8-OHdG) (-52%, -44%) compared with the WRF or HC diets (P < 0.05). There were no differences in aortic 8-OHdG levels between rabbits fed the BRF and normal diets. The BRF diet significantly (P < 0.05) decreased the malondialdehyde (MDA) level of serum (-37%) and aortic artery (-50%) compared with the WRF diet. There were no differences in the concentrations of serum total cholesterol (TC), LDL cholesterol (LDL-C), HDL-C or the ratio of apoprotein (apo)I/apoB among the HC, WRF and BRF groups. Similarly, there were no differences in the serum vitamin E concentration and erythrocyte and aorta superoxide dismutase (SOD) activities among rabbits fed these diets. The serum concentration of most fatty acids except 18:1 did not differ between the WRF and the BRF groups. We conclude that the inhibition of atherosclerotic plaque formation derived from the black rice outer layer fraction in rabbits might be mediated by antioxidative or anti-inflammatory effects. PMID:11773502

  19. Oritavancin Pharmacokinetics and Bone Penetration in Rabbits

    PubMed Central

    Ostiguy, Valerie; Cadieux, Cordelia; Malouin, Mireille; Belanger, Odette; Far, Adel Rafai; Parr, Thomas R.

    2015-01-01

    The pharmacokinetics and bone concentrations of oritavancin were investigated after a single intravenous dose was administered to rabbits. The pharmacokinetic profile of oritavancin in rabbits showed that it is rapidly distributed to bone tissues, with concentrations remaining stable for up to 168 h, the last measured time point. Based on these findings, further evaluation of oritavancin for the treatment of infections in bone tissues is warranted. PMID:26239977

  20. Erythrocytes in nanomedicine: an optimal blend of natural and synthetic materials.

    PubMed

    Zhang, Haijun

    2016-07-21

    A hybrid approach strategy using synthetic nanoparticles and erythrocytes offers an optimal blend of natural and synthetic materials. The combined advantages of erythrocytes and nanoparticles could serve as an immune-evasive multifunctional platform. This review summarized the research on state-of-the-art and significant advances in erythrocytes for nanomedicine, and presented are their fabrication process, their unique properties and applications. According to its structure, this review mainly focuses on three kinds of erythrocyte-based nanomedicine: whole erythrocytes as carriers, cell membrane coated nanoparticles, and nanoerythrosomes. In addition, some future prospects are also prudentially addressed. We expect rapid success in the advancement of erythrocyte-based nanomedicines, from the bench to the bedside.

  1. Anti-Self Phosphatidylserine Antibodies Recognize Uninfected Erythrocytes Promoting Malarial Anemia.

    PubMed

    Fernandez-Arias, Cristina; Rivera-Correa, Juan; Gallego-Delgado, Julio; Rudlaff, Rachel; Fernandez, Clemente; Roussel, Camille; Götz, Anton; Gonzalez, Sandra; Mohanty, Akshaya; Mohanty, Sanjib; Wassmer, Samuel; Buffet, Pierre; Ndour, Papa Alioune; Rodriguez, Ana

    2016-02-10

    Plasmodium species, the parasitic agents of malaria, invade erythrocytes to reproduce, resulting in erythrocyte loss. However, a greater loss is caused by the elimination of uninfected erythrocytes, sometimes long after infection has been cleared. Using a mouse model, we found that Plasmodium infection induces the generation of anti-self antibodies that bind to the surface of uninfected erythrocytes from infected, but not uninfected, mice. These antibodies recognize phosphatidylserine, which is exposed on the surface of a fraction of uninfected erythrocytes during malaria. We find that phosphatidylserine-exposing erythrocytes are reticulocytes expressing high levels of CD47, a "do-not-eat-me" signal, but the binding of anti-phosphatidylserine antibodies mediates their phagocytosis, contributing to anemia. In human patients with late postmalarial anemia, we found a strong inverse correlation between the levels of anti-phosphatidylserine antibodies and plasma hemoglobin, suggesting a similar role in humans. Inhibition of this pathway may be exploited for treating malarial anemia.

  2. Modifications of a cholinesterase method for determination of erythrocyte cholinesterase activity in wild mammals.

    PubMed

    Donovan, D A; Zinkl, J G

    1994-04-01

    A method to determine erythrocyte cholinesterase (ChE) activity was modified for use in wild mammals. Erythrocyte ChE of California voles (Microtus californicus) was primarily acetylcholinesterase (AChE), which was similar to the brain and unlike plasma which was primarily butyrylcholinesterase (BChE). Triplicate erythrocyte AChE analyses from individual animals of several species of wild rodents revealed a mean coefficient of variation of 8.7% (SD = 4.3%). Erythrocyte ChE activity of several wild mammals of California revealed that mule deer (Odocoileus hemionus) had the highest erythrocyte AChE activity (1,514.5 mU/ml) and dusky-footed woodrats (Neotoma fuscipes) had the lowest activity (524.3 mU/ml). No ChE activity was found in erythrocytes of several species of birds and fish. PMID:8028108

  3. Reverse-Transcriptase PCR Detection of Leptospira: Absence of Agreement with Single-Specimen Microscopic Agglutination Testing

    PubMed Central

    Waggoner, Jesse J.; Balassiano, Ilana; Mohamed-Hadley, Alisha; Vital-Brazil, Juliana Magalhães; Sahoo, Malaya K.; Pinsky, Benjamin A.

    2015-01-01

    Background Reference diagnostic tests for leptospirosis include nucleic acid amplification tests, bacterial culture, and microscopic agglutination testing (MAT) of acute and convalescent serum. However, clinical laboratories often do not receive paired specimens. In the current study, we tested serum samples using a highly sensitive real-time nucleic acid amplification test for Leptospira and compared results to MAT performed on the same specimens. Methods/Principal Findings 478 serum samples from suspected leptospirosis cases in Rio de Janeiro were tested using a real-time RT-PCR for the diagnosis of leptospirosis, malaria and dengue (the Lepto-MD assay). The Lepto-MD assay detects all species of Leptospira (saprophytic, intermediate, and pathogenic), and in the current study, we demonstrate that this assay amplifies both Leptospira RNA and DNA. Dengue virus RNA was identified in 10 patients, and no cases of malaria were detected. A total of 65 samples (13.6%) were positive for Leptospira: 35 samples (7.3%) in the Lepto-MD assay, 33 samples (6.9%) by MAT, and 3 samples tested positive by both (kappa statistic 0.02). Poor agreement between methods was consistent regardless of the titer used to define positive MAT results or the day of disease at sample collection. Leptospira nucleic acids were detected in the Lepto-MD assay as late as day 22, and cycle threshold values did not differ based on the day of disease. When Lepto-MD assay results were added to the MAT results for all patients in 2008 (n=818), the number of detected leptospirosis cases increased by 30.4%, from 102 (12.5%) to 133 (16.3%). Conclusions/Significance This study demonstrates a lack of agreement between nucleic acid detection of Leptospira and single-specimen MAT, which may result from the clearance of bacteremia coinciding with the appearance of agglutinating antibodies. A combined testing strategy for acute leptospirosis, including molecular and serologic testing, appears necessary to maximize

  4. 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

  5. Genomic Analysis of Companion Rabbit Staphylococcus aureus.

    PubMed

    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

  6. Genomic Analysis of Companion Rabbit Staphylococcus aureus.

    PubMed

    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.

  7. Functional significance of glutamate-cysteine ligase modifier for erythrocyte survival in vitro and in vivo.

    PubMed

    Föller, M; Harris, I S; Elia, A; John, R; Lang, F; Kavanagh, T J; Mak, T W

    2013-10-01

    Erythrocytes endure constant exposure to oxidative stress. The major oxidative stress scavenger in erythrocytes is glutathione. The rate-limiting enzyme for glutathione synthesis is glutamate-cysteine ligase, which consists of a catalytic subunit (GCLC) and a modifier subunit (GCLM). Here, we examined erythrocyte survival in GCLM-deficient (gclm(-/-)) mice. Erythrocytes from gclm(-/-) mice showed greatly reduced intracellular glutathione. Prolonged incubation resulted in complete lysis of gclm(-/-) erythrocytes, which could be reversed by exogenous delivery of the antioxidant Trolox. To test the importance of GCLM in vivo, mice were treated with phenylhydrazine (PHZ; 0.07 mg/g b.w.) to induce oxidative stress. Gclm(-/-) mice showed dramatically increased hemolysis compared with gclm(+/+) controls. In addition, PHZ-treated gclm(-/-) mice displayed markedly larger accumulations of injured erythrocytes in the spleen than gclm(+/+) mice within 24 h of treatment. Iron staining indicated precipitations of the erythrocyte-derived pigment hemosiderin in kidney tubules of gclm(-/-) mice and none in gclm(+/+) controls. In fact, 24 h after treatment, kidney function began to diminish in gclm(-/-) mice as evident from increased serum creatinine and urea. Consequently, while all PHZ-treated gclm(+/+) mice survived, 90% of PHZ-treated gclm(-/-) mice died within 5 days of treatment. In vitro, upon incubation in the absence or presence of additional oxidative stress, gclm(-/-) erythrocytes exposed significantly more phosphatidylserine, a cell death marker, than gclm(+/+) erythrocytes, an effect at least partially due to increased cytosolic Ca(2+) concentration. Under resting conditions, gclm(-/-) mice exhibited reticulocytosis, indicating that the enhanced erythrocyte death was offset by accelerated erythrocyte generation. GCLM is thus indispensable for erythrocyte survival, in vitro and in vivo, during oxidative stress.

  8. Parathyroid hormone ablation alters erythrocyte parameters that are rescued by calcium-sensing receptor gene deletion.

    PubMed

    Romero, Jose R; Youte, Rodeler; Brown, Edward M; Pollak, Martin R; Goltzman, David; Karaplis, Andrew; Pong, Lie-Chin; Chien, Lawrence; Chattopadhyay, Naibedya; Rivera, Alicia

    2013-07-01

    The mechanisms by which parathyroid hormone (PTH) produces anemia are unclear. Parathyroid hormone secretion is regulated by the extracellular Ca2+ -sensing receptor. We investigated the effects of ablating PTH on hematological indices and erythrocytes volume regulation in wild-type, PTH-null, and Ca2+ -sensing receptor-null/PTH-null mice. The erythrocyte parameters were measured in whole mouse blood, and volume regulatory systems were determined by plasma membrane K+ fluxes, and osmotic fragility was measured by hemoglobin determination at varying osmolarities. We observed that the absence of PTH significantly increases mean erythrocyte volume and reticulocyte counts, while decreasing erythrocyte counts, hemoglobin, hematocrit, and mean corpuscular hemoglobin concentration. These changes were accompanied by increases in erythrocyte cation content, a denser cell population, and increased K+ permeability, which were in part mediated by activation of the K+ /Cl- cotransporter and Gardos channel. In addition we observed that erythrocyte osmotic fragility in PTH-null compared with wild-type mice was enhanced. When Ca2+ -sensing receptor gene was deleted on the background of PTH-null mice, we observed that several of the alterations in erythrocyte parameters of PTH-null mice were largely rescued, particularly those related to erythrocyte volume, K+ fluxes and osmotic fragility, and became similar to those observed in wild-type mice. Our results demonstrate that Ca2+ -sensing receptor and parathyroid hormone are functionally coupled to maintain erythrocyte homeostasis. PMID:23528155

  9. Changes in the nuclear matrix of chicken erythrocytes that accompany maturation.

    PubMed Central

    Chen, H Y; Sun, J M; Hendzel, M J; Rattner, J B; Davie, J R

    1996-01-01

    The protein composition and structure of nuclear matrices isolated from adult chicken immature and mature erythrocytes were analysed. Visualization of nuclear matrices by electron microscopy showed that immature-erythrocyte nuclear matrices had internal structures, while most mature-erythrocyte nuclear matrices did not. Both mature- and immature-erythrocyte nuclear matrices were surrounded by a fibrous network of intermediate filaments. Two-dimensional gel electrophoretic analysis of proteins obtained from fractionated nuclear matrices led to the assignment of the proteins as components of the nuclear porelamina, internal matrix, or cytoskeleton. Common and different proteins belonging to one of the three groups were identified in nuclear matrices of immature and mature erythrocytes. Investigation of the partitioning of histone deacetylase activity, an enzyme associated with the internal matrix, among the erythroid nuclear matrix fractions provided evidence that mature- and immature-erythrocyte nuclear matrices have internal structures. However, the activity of histone deacetylase and level of internal matrix proteins from mature-erythrocyte nuclear matrices were less than those from immature-erythrocyte matrices. The low levels of nuclear RNA and internal matrix proteins may account for lack of visual evidence for an internal matrix in mature erythrocytes. PMID:8947496

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

    SciTech Connect

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

    1987-12-01

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

  11. Aggregation ability of erythrocytes of patients with coronary heart disease depending on different glucose concentration

    NASA Astrophysics Data System (ADS)

    Malinova, Lidia I.; Simonenko, Georgy V.; Kirichuk, Vyacheslav F.; Denisova, Tatyana P.; Tuchin, Valery V.

    2002-07-01

    The aggregation ability of erythrocytes of patients with coronary heart disease comparing to practically healthy persons and patients with coronary heart disease combined with non insulin dependent diabetes mellitus depending on different glucose concentration in unguentums of blood incubates with the help of computer microphotometer - visual analyzer was studied. Two-phase behavior of erythrocytes size changing of practically healthy persons depending on glucose concentration in an incubation medium and instability erythrocyte systems of a whole blood to the influence of high glucose concentration were revealed. Influence of high glucose concentration on aggregation ability of erythrocytes of patients with coronary heart disease and its combination with non insulin dependent diabetes mellitus was revealed.

  12. Insect erythrocyte agglutinins. In vitro opsonization experiments with Clitumnus extradentatus and Periplaneta americana haemocytes.

    PubMed Central

    Rowley, A F; Ratcliffe, N A

    1980-01-01

    The effect of naturally occurring haemagglutinins on the in vitro phagocytosis of sheep erythrocytes by the blood cells (haemocytes) of Clitumnus extradentatus and Periplaneta americana was studied. The results showed that the haemagglutinins in both species failed to act as opsonins. Indeed, in some instances, incubation of erythrocytes in haemolymph resulted in less avid ingestion as compared with the saline-incubated controls. This reduced phagocytosis was probably caused by the clumping of erythrocytes on the haemocyte monolayers, leaving fewer single red cells available for uptake. The possible roles of these erythrocyte agglutinins in the host defence systems of insects are discussed. Images Figures 1-3 Figures 6-7 PMID:7000682

  13. A Lectin-Like Receptor is Involved in Invasion of Erythrocytes by Plasmodium falciparum

    NASA Astrophysics Data System (ADS)

    Jungery, M.; Pasvol, G.; Newbold, C. I.; Weatherall, D. J.

    1983-02-01

    Glycophorin both in solution and inserted into liposomes blocks invasion of erythrocytes by the malaria parasite Plasmodium falciparum. Furthermore, one sugar, N-acetyl-D-glucosamine (GlcNAc), completely blocks invasion of the erythrocyte by this parasite. GlcNAc coupled to bovine serum albumin to prevent the sugar entering infected erythrocytes was at least 100,000 times more effective than GlcNAc alone. Bovine serum albumin coupled to lactose or bovine serum albumin alone had no effect on invasion. These results suggest that the binding of P. falciparum to erythrocytes is lectin-like and is determined by carbohydrates on glycophorin.

  14. Differences in the number of micronucleated erythrocytes among young and adult animals including humans. Spontaneous micronuclei in 43 species.

    PubMed

    Zúñiga-González, G; Torres-Bugarín, O; Zamora-Perez, A; Gómez-Meda, B C; Ramos Ibarra, M L; Martínez-González, S; González-Rodríguez, A; Luna-Aguirre, J; Ramos-Mora, A; Ontiveros-Lira, D; Gallegos-Arreola, M P

    2001-07-25

    In our previous report we speculated about the possibility that some species had high levels of spontaneous micronucleated erythrocytes (MNE) just in a juvenile stage, this is, that the MNE diminish as the reticuloendothelial system matures. Here we show this effect in species including rat, rabbit, pig, dog, cat, gray squirrel, lion, giraffe, white-tailed deer, opossum and even human. The number of spontaneous MNE that we found in 43 species is shown, and the proportions of polychromatic and normochromatic. This is our third report on spontaneous MNE in different species. We obtained 189 peripheral blood samples of mammals, birds and reptiles. From 12 species we obtained only one sample, and 16 were reported previously, but now the size of the sample has been increased. The species with the highest spontaneous MNE were the Vietnamese potbelly pig (with the highest MNE number), Bengal tiger, capuchin monkey, puma, ferret, owl, hedgehog, squirrel monkey, pig and white-tailed deer. These species could be used as monitors for genotoxic events.

  15. Differences in the number of micronucleated erythrocytes among young and adult animals including humans. Spontaneous micronuclei in 43 species.

    PubMed

    Zúñiga-González, G; Torres-Bugarín, O; Zamora-Perez, A; Gómez-Meda, B C; Ramos Ibarra, M L; Martínez-González, S; González-Rodríguez, A; Luna-Aguirre, J; Ramos-Mora, A; Ontiveros-Lira, D; Gallegos-Arreola, M P

    2001-07-25

    In our previous report we speculated about the possibility that some species had high levels of spontaneous micronucleated erythrocytes (MNE) just in a juvenile stage, this is, that the MNE diminish as the reticuloendothelial system matures. Here we show this effect in species including rat, rabbit, pig, dog, cat, gray squirrel, lion, giraffe, white-tailed deer, opossum and even human. The number of spontaneous MNE that we found in 43 species is shown, and the proportions of polychromatic and normochromatic. This is our third report on spontaneous MNE in different species. We obtained 189 peripheral blood samples of mammals, birds and reptiles. From 12 species we obtained only one sample, and 16 were reported previously, but now the size of the sample has been increased. The species with the highest spontaneous MNE were the Vietnamese potbelly pig (with the highest MNE number), Bengal tiger, capuchin monkey, puma, ferret, owl, hedgehog, squirrel monkey, pig and white-tailed deer. These species could be used as monitors for genotoxic events. PMID:11423355

  16. ORAL PAPILLOMATOSIS OF RABBITS: A VIRUS DISEASE

    PubMed Central

    Parsons, Robert J.; Kidd, John G.

    1943-01-01

    Papillomas occur frequently on the oral mucosa of domestic rabbits procured in the metropolitan area of New York. They are small and benign, and are situated mostly on the under side of the tongue. A filtrable virus can be extracted from them with which growths can be reproduced in the oral mucosa of several species of rabbits and hares but which fails to cause lesions when inoculated into other rabbit tissues and into the oral mucosa of other species. The virus differs notably from the Shope virus, which causes cutaneous papillomas in rabbits but proves innocuous to oral mucosa: rabbits solidly immune to the oral papilloma virus are fully susceptible to the Shope virus and vice versa. The oral papillomas are not highly contagious, for susceptible animals kept in individual cages in the same rooms with others carrying the growths, fed the same kind of food, and cared for by the same attendants, do not "catch" them. They are found much more frequently in the offspring of dams that carry the growths than in those of mothers free from them, and the causative virus can be recovered from the mouth washings of rabbits having no growths. The observations indicate that the virus may be spread by transfer from the mother to the young during the period of suckling, and that it may lie latent in the mouth, doing no harm unless the mucous membrane is injured. The slight trauma occurring now and then when coarse foods are chewed may furnish the required tissue nidus under natural conditions, for papillomas occasionally appear after virus has been dropped into the mouths of uninoculated rabbits; but the more extensive injury and healing resulting from experimental tattoo inoculations proves regularly effective in this respect. Tar can also act as an efficient adjuvant to the virus, the incidence of "spontaneous" oral papillomas being much higher in domestic rabbits that had had the opportunity to lick tar from their ears and paws during long periods than in normal control

  17. Collaborative evaluation of antigen detection by a commercial latex agglutination test and enzyme immunoassay in the diagnosis of invasive candidiasis.

    PubMed Central

    Lemieux, C; St-Germain, G; Vincelette, J; Kaufman, L; de Repentigny, L

    1990-01-01

    The Cand-Tec Candida detection system and enzyme immunoassay for serum mannan were retrospectively compared in a controlled collaborative evaluation of antigen detection in 32 patients with candidiasis proven by biopsy or culture from a normally sterile site and with sera drawn within 7 days of inclusion. With a threshold titer of 1/8, which excluded false-positive results in 17 hospitalized patients without candidiasis, sensitivities for all 32 patients with candidiasis were 44% for the Cand-Tec assay and 17% for the enzyme immunoassay. Both assays provided greater sensitivity when sera were drawn within 24 h of inclusion in the study and in the category of patients with invasive candidiasis (57% by Cand-Tec and 33% by enzyme immunoassay). The Cand-Tec assay gave false-positive results (titer, greater than or equal to 1/8) in 4 of 6 patients with transient candidemia, in 1 of 20 otherwise healthy patients with rheumatoid factor, and in 1 patient with a positive cryptococcal latex agglutination test. Three serum specimens from 3 of 32 patients with candidiasis contained rheumatoid factor and gave titers of greater than or equal to 1/8 by the Cand-Tec assay. Detection of serum mannan by enzyme immunoassay was less sensitive but more specific than the Cand-Tec Candida detection system for the diagnosis of invasive candidiasis. PMID:2179258

  18. Comparative evaluation of Rose Bengal plate agglutination test, mallein test, and some conventional serological tests for diagnosis of equine glanders.

    PubMed

    Naureen, Abeera; Saqib, Muhammad; Muhammad, Ghulan; Hussain, Muhammad H; Asi, Muhammad N

    2007-07-01

    The Rose Bengal plate agglutination test (RBT) was evaluated for the diagnosis of equine glanders, and its diagnostic efficiency was compared with that of mallein and other serological tests, including indirect hemagglutination test (IHAT), complement fixation test (CFT), and modified counter immunoelectrophoresis test (mCIET). Sera from 70 naturally infected culture-positive, 96 potentially exposed cohorts, and 110 healthy equines were tested. All tests but mCIET showed 100% specificity when testing the sera from glanders-negative equines. The calculated sensitivities of RBT, IHAT, CFT, mCIET, and mallein test when testing culture-positive equines were 90.0, 97.1, 91.4, 81.4, and 75.7%, respectively. The RBT was significantly (P < 0.05) more sensitive than the mallein test and mCIET. The positive and negative predictive values of each test (RBT, IHAT, CFT, mallein test, and mCIET) were as follows: 100 and 94, 100 and 98.2, 100 and 96.7, 100 and 86.6, and 90.5 and 88.6, respectively. On comparing glandered and nonglandered animals, the highest agreement (0.987) was found between RBT and CFT followed by RBT and IHAT (0.940), RBT and mallein test (0.871), and RBT and mCIET (0.852). Because the RBT is simpler and rapid to perform, the inclusion of the test as a supplementary test for the diagnosis of glanders in field conditions is recommended.

  19. Sero-epidemiology of equine toxoplasmosis using a latex agglutination test in the three metropolises of Punjab, Pakistan.

    PubMed

    Saqib, M; Hussain, M H; Sajid, M S; Mansoor, M K; Asi, M N; Fadya, A A K; Zohaib, A; Sial, A U R; Muhammad, G; Ullah, I

    2015-06-01

    Toxoplasmosis is a serious threat for livestock in addition to being of zoonotic significance. In this study, serodiagnosis of equine toxoplasmosis was conducted in a randomly selected population from the 3 metropolises of Punjab, Pakistan. To this end, 272 draught equines were screened using a commercial latex agglutination assay kit. Association of probable risk factors of equine toxoplasmosis was also documented. A total of 91 (33.5%) equines were found sero-positive for Toxoplama (T.) gondii having antibody titers ranging between 1:32 to 1:612. The highest rates of seropositive cases were observed in donkeys (58.7%) followed by mules (28.6%) and horses (23.5%). Age, sex and species of draught equines were found not to be statistically (p>0.05) associated with the distribution of T. gondii antibodies. The results of the study provided a baseline data for the exposure of equine population in this area. In addition, it is recommended that the contiguous population of domestic ruminants and possible reservoirs such as feral cats should be screened in order to explore the potential risk for the human population in Pakistan.

  20. Comparison of a dot immunobinding assay, latex agglutination, and cytotoxin assay for laboratory diagnosis of Clostridium difficile-associated diarrhea.

    PubMed

    Woods, G L; Iwen, P C

    1990-05-01

    C. diff-CUBE, a dot immunobinding assay (DIA) (Difco Laboratories, Ann Arbor, Mich.) for detection of Clostridium difficile toxin A in stool specimens, was compared with latex agglutination (LA) (Marion Laboratories, Kansas City, Mo.) and cytotoxin assay (CTA) for the laboratory diagnosis of C. difficile-associated diarrhea. A total of 200 stool specimens collected from 169 patients with suspected C. difficile diarrhea were tested. Of the 198 specimens evaluated by all three methods, 36 (18%) from 36 patients were positive by one or more of the tests. Twenty-five, 26, and 23 specimens were positive by CTA, DIA, and LA, respectively; 14 were positive by all three methods. Eight specimens yielded nonspecific LA test results; all eight were negative by CTA, and one was positive by DIA. DIA results agreed with CTA results in 183 (92%) cases and with LA results in 175 (88%) cases. CTA and LA results agreed in 179 (90%) cases. Freezing of the specimen did not appear to adversely affect either the DIA or LA test. These preliminary results suggest that C. diff-CUBE may be useful as a rapid screen for the diagnosis of C. difficile-associated diarrhea. However, for optimum laboratory diagnosis, further testing of all stools that are negative by DIA is warranted.

  1. Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test.

    PubMed Central

    Fach, P; Popoff, M R

    1997-01-01

    A duplex PCR procedure was evaluated for the detection of Clostridium perfringens in food and biological samples and for the identification of enterotoxigenic strains. This method uses two sets of primers which amplify in the same reaction two different DNA fragments simultaneously: the 283-bp C. perfringens phospholipase C gene fragment and the 426-bp enterotoxin gene fragment. Internal primers within the two primer sets confirmed the specificity of the method by DNA-DNA hybridization with the PCR products. No cross-reaction was observed with other Clostridium species or with other bacteria routinely found in food. The detection level was approximately 10(5) C. perfringens cells per g of stool or food sample. When overnight enrichment culture was used, 10 C. perfringens cells per g was detected in 57 artificially contaminated food samples. The duplex PCR is a rapid, sensitive, and reliable method for the detection and identification of enterotoxigenic C. perfringens strains in food samples. A slide latex agglutination test was also evaluated as a rapid, simple technique for the detection of C. perfringens enterotoxin in stool samples. PMID:9361409

  2. A zebrafish intelectin ortholog agglutinates both Gram-negative and Gram-positive bacteria with binding capacity to bacterial polysaccharide.

    PubMed

    Chen, Lei; Yan, Jie; Sun, Weiping; Zhang, Yan; Sui, Chao; Qi, Jing; Du, Yijun; Feng, Lijun

    2016-08-01

    Intelectins are glycan-binding lectins found in various species including cephalochordates, urochordates, fish, amphibians and mammals. But their detailed functions are not well studied in zebrafish which is a good model to study native immunity. In this study, we cloned a zebrafish intelectin ortholog, zebrafish intelectin 2 (zITLN2), which contains a conserved fibrinogen-related domain (FReD) in the N-terminus and the unique intelectin domain in the C-terminus. We examined the tissue distribution of zITLN2 in adult zebrafish and found that zITLN2 was expressed in various organs with the highest level in intestine. Like amphioxus intelectins, zITLN2 expression was upregulated in adult zebrafish infected with Staphylococcus aureus with the highest expression level at 12 h after challenge. Recombinant zITLN2 protein expressed in E. coli was able to agglutinate both Gram-negative and Gram-positive bacteria to similar degrees in a calcium-dependent manner. Furthermore, recombinant zITLN2 bound lipopolysaccharide (LPS) and peptidoglycan (PGN) comparably. Our work on zITLN2 provided further information to understand functions of this new family of lectins and the innate immunity in vertebrates. PMID:27329687

  3. Legionella species and serogroups in Malaysian water cooling towers: identification by latex agglutination and PCR-DNA sequencing of isolates.

    PubMed

    Yong, Stacey Foong Yee; Goh, Fen-Ning; Ngeow, Yun Fong

    2010-03-01

    In this study, we investigated the distribution of Legionella species in water cooling towers located in different parts of Malaysia to obtain information that may inform public health policies for the prevention of legionellosis. A total of 20 water samples were collected from 11 cooling towers located in three different states in east, west and south Malaysia. The samples were concentrated by filtration and treated with an acid buffer before plating on to BCYE agar. Legionella viable counts in these samples ranged from 100 to 2,000 CFU ml(-1); 28 isolates from the 24 samples were examined by latex agglutination as well as 16S rRNA and rpoB PCR-DNA sequencing. These isolates were identified as Legionella pneumophila serogroup 1 (35.7%), L. pneumophila serogroup 2-14 (39%), L. pneumophila non-groupable (10.7%), L. busanensis, L. gormanii, L. anisa and L. gresilensis. L. pneumophila was clearly the predominant species at all sampling sites. Repeat sampling from the same cooling tower and testing different colonies from the same water sample showed concurrent colonization by different serogroups and different species of Legionella in some of the cooling towers. PMID:20009251

  4. Kinetics of acute inflammation induced by Escherichia coli in rabbits. II. The effect of hyperimmunization, complement depletion, and depletion of leukocytes.

    PubMed Central

    Kopaniak, M. M.; Movat, H. Z.

    1983-01-01

    The inflammatory response to Escherichia coli was quantitated in the skin of normal rabbits and the kinetics established as described previously. Hyperemia, measured with radiolabeled microspheres; vascular permeability, estimated with 125 I-albumin; and leukocyte infiltration, quantitated with 51Cr-labeled autologous leukocytes, reached maximal values 3 hours after the injection of bacteria and subsided almost completely by 6 hours. Hemorrhage, measured with homologous 59Fe-erythrocytes, continued to increase between 1 and 6 hours after injection and then reached plateau levels. The lesions were studied up to 8 hours, since in the previous study no changes were observed beyond that time. In the study described in this paper, the host mediation systems were manipulated in various groups of rabbits in order to elucidate the mechanisms underlying the development of the inflammatory reaction. One group of animals was hyperimmunized with the E coli organisms, another was partially depleted of hemolytic complement with cobra venom factor, and yet another was rendered leukopenic with nitrogen mustard. In hyperimmunized animals hyperemia in the dermal lesions induced by the microorganisms was significantly more intense than in normal rabbits. Vascular permeability increase occurred earlier in hyperimmunized rabbits and at 1 hour was significantly greater than in normals. Decomplemented rabbits had significantly less vascular permeability than normal animals, whereas in leukopenic rabbits no increase in vascular permeability could be elicited. Leukocyte accumulation was increased over the normal animals in the lesions of hyperimmunized rabbits. Hemorrhage was significantly decreased in leukopenic rabbits. Histologic examination of the lesions revealed that whereas in normal animals the infiltrating neutrophils ingested most of the bacteria and formed definite abscesses by 6-8 hours, these abscesses were absent in leukopenic animals, and free-lying bacteria were

  5. Phosphatidylinositol kinase from rabbit reticulocytes

    SciTech Connect

    Tuazon, P.T.; Heng, A.B.W.; Traugh, J.A.

    1986-05-01

    Phosphatidylinositol (PI) kinase was isolated from the postribosomal supernatant of rabbit reticulocytes. This activity was identified by the formation of a product that comigrated with phosphatidylinositol-4-phosphate (PIP) when purified PI was phosphorylated in the presence of (/sup 32/P)ATP and Mg/sup 2 +/. Three major peaks of PI kinase activity were resolved by chromatography on DEAE-cellulose. The first peak eluted at 50-100 mM NaCl together with several serine protein kinases, casein kinase (CK) I and protease activated kinase (PAK) I and II. The PI kinase was subsequently separated from the protein kinases by chromatography on phosphocellulose. The second peak eluted at 125-160 mM NaCl and contained another lipid kinase activity that produced a product which comigrated with phosphatidic acid on thin layer chromatography. The third peak, which eluted at 165-200 mM NaCl, partly comigrated with casein kinase (CK) II and an active protein kinase(s) which phosphorylated mixed histone and histone I. CK II and the histone kinase activities were also separated by chromatography on phosphocelluslose. The different forms of PI kinase were characterized and compared with respect to substrate and salt requirements.

  6. Ultrasonic Backscattering from Suspended Erythrocytes: Dependence on Frequency and Size.

    NASA Astrophysics Data System (ADS)

    Kuo, Ihyuan

    The ultrasonic scattering properties of blood have been intensively investigated since the echo signal from red blood cells carries abundant diagnostic information for the study of blood flow and blood properties in the vessels. Recently, ultrasound of frequency higher than 20 MHz has been implemented in intravascular imaging to obtain better images of the vessel wall. In this research measurements were extended to 30 MHz to better understand the effect of blood on the operation of these intravascular devices. The experimentally measured backscatter of saline suspended porcine erythrocytes for frequency up to 30 MHz agrees very well with the theoretical analysis which indicate that Rayleigh scattering is still valid below this frequency. The analysis utilize the T-matrix method to calculate the backscattering cross section of an erythrocyte modeled as a fluid sphere, disk, and biconcave disk. Measurements on the backscattering coefficients of porcine, bovine, and lamb erythrocytes reveal that the backscatter has a square dependence on cell volume. The cell size dependent backscatter is also analyzed via a continuum approach. It is found that the echo intensity of high frequency ultrasound suffers greatly from the attenuation. The dilemma may be solved by using a spherically focused transducer. An analysis of the focused beam reflected from a perfect planar reflector leads to the modification of the standard substitution method for the backscatter measurement since the "image source" theory is found to be inappropriate for the focused beam. Reflection of the focused beam near the focal point is described based on Huygens' principle. Experimental and theoretical results indicate that the backscatter is dependent upon the position of the scatterer and the geometry of the transducer if a focused beam is used. Since ultrasound velocity information is needed for scattering measurements, an innovative method for measuring the acoustic speed and the attenuation coefficient

  7. Human erythrocytes analyzed by generalized 2D Raman correlation spectroscopy

    NASA Astrophysics Data System (ADS)

    Wesełucha-Birczyńska, Aleksandra; Kozicki, Mateusz; Czepiel, Jacek; Łabanowska, Maria; Nowak, Piotr; Kowalczyk, Grzegorz; Kurdziel, Magdalena; Birczyńska, Malwina; Biesiada, Grażyna; Mach, Tomasz; Garlicki, Aleksander

    2014-07-01

    The most numerous elements of the blood cells, erythrocytes, consist mainly of two components: homogeneous interior filled with hemoglobin and closure which is the cell membrane. To gain insight into their specific properties we studied the process of disintegration, considering these two constituents, and comparing the natural aging process of human healthy blood cells. MicroRaman spectra of hemoglobin within the single RBC were recorded using 514.5, and 785 nm laser lines. The generalized 2D correlation method was applied to analyze the collected spectra. The time passed from blood donation was regarded as an external perturbation. The time was no more than 40 days according to the current storage limit of blood banks, although, the average RBC life span is 120 days. An analysis of the prominent synchronous and asynchronous cross peaks allow us to get insight into the mechanism of hemoglobin decomposition. Appearing asynchronous cross-peaks point towards globin and heme separation from each other, while synchronous shows already broken globin into individual amino acids. Raman scattering analysis of hemoglobin “wrapping”, i.e. healthy erythrocyte ghosts, allows for the following peculiarity of their behavior. The increasing power of the excitation laser induced alterations in the assemblage of membrane lipids. 2D correlation maps, obtained with increasing laser power recognized as an external perturbation, allows for the consideration of alterations in the erythrocyte membrane structure and composition, which occurs first in the proteins. Cross-peaks were observed indicating an asynchronous correlation between the senescent-cell antigen (SCA) and heme or proteins vibrations. The EPR spectra of the whole blood was analyzed regarding time as an external stimulus. The 2D correlation spectra points towards participation of the selected metal ion centers in the disintegration process.

  8. Rapid Effects of a Protective O-Polysaccharide-Specific Monoclonal IgA on Vibrio cholerae Agglutination, Motility, and Surface Morphology

    PubMed Central

    Levinson, Kara J.; De Jesus, Magdia

    2015-01-01

    2D6 is a dimeric monoclonal immunoglobulin A (IgA) specific for the nonreducing terminal residue of Ogawa O-polysaccharide (OPS) of Vibrio cholerae. It was previously demonstrated that 2D6 IgA is sufficient to passively protect suckling mice from oral challenge with virulent V. cholerae O395. In this study, we sought to define the mechanism by which 2D6 IgA antibody protects the intestinal epithelium from V. cholerae infection. In a mouse ligated-ileal-loop assay, 2D6 IgA promoted V. cholerae agglutination in the intestinal lumen and limited the ability of the bacteria to associate with the epithelium, particularly within the crypt regions. In vitro fluorescence digital video microscopy analysis of antibody-treated V. cholerae in liquid medium revealed that 2D6 IgA not only induced the rapid (5- to 10-min) onset of agglutination but was an equally potent inhibitor of bacterial motility. Scanning electron microscopy showed that 2D6 IgA promoted flagellum-flagellum cross-linking, as well as flagellar entanglement with bacterial bodies, suggesting that motility arrest may be a consequence of flagellar tethering. However, monovalent 2D6 Fab fragments also inhibited V. cholerae motility, demonstrating that antibody-mediated agglutination and motility arrest are separate phenomena. While 2D6 IgA is neither bactericidal nor bacteriostatic, exposure of V. cholerae to 2D6 IgA (or Fab fragments) resulted in a 5-fold increase in surface-associated blebs, as well an onset of a wrinkled surface morphotype. We propose that the protective immunity conferred by 2D6 IgA is the result of multifactorial effects on V. cholerae, including agglutination, motility arrest, and possibly outer membrane stress. PMID:25667263

  9. Rapid Detection of Contagious Caprine Pleuropneumonia Using a Mycoplasma capricolum subsp. capripneumoniae Capsular Polysaccharide-Specific Antigen Detection Latex Agglutination Test

    PubMed Central

    March, J. B.; Gammack, C.; Nicholas, R.

    2000-01-01

    Latex microspheres (diameter, 8 μm) were coated with anti-Mycoplasma capricolum subsp. capripneumoniae polyclonal immunoglobulin G (IgG) antiserum (anti-F38 biotype). The coated microspheres, when used in a latex agglutination test (LAT), detected M. capricolum subsp. capripneumoniae antigen in the serum of goats with contagious caprine pleuropneumoniae (CCPP). Beads also agglutinated strongly in the presence of purified M. capricolum subsp. capripneumoniae capsular polysaccharide (CPS). Preabsorption of CPS-specific antibodies prior to coating of the beads removed agglutinating activity in the presence of M. capricolum subsp. capripneumoniae, strongly suggesting that CPS is the likely soluble antigen recognized by the test. In addition, the specificity of the LAT exactly mirrored that of an M. capricolum subsp. capripneumoniae CPS-specific monoclonal antibody (WM25): of the 8 other mycoplasma species tested, agglutination was observed only with bovine serogroup 7. The LAT detected all 11 strains of M. capricolum subsp. capripneumoniae examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 1.7 × 104 CFU, in a reaction volume of 0.03 ml of serum. With field sera from goats with CCPP, the results of the LAT exhibited a 67% correlation with the results of the currently used complement fixation test (CFT), with the main discrepancy in diagnosis resulting from the increased sensitivity of the LAT compared to that of CFT. This antigen-detection LAT should prove particularly useful in identifying animals in the earliest stages of CCPP and combines sensitivity and low cost with ease of application in the field, without the need for any specialist training or equipment. PMID:11060083

  10. Prevalence of agglutinating antibodies to Toxoplasma gondii in striped skunks (Mephitis mephitis), opossums (Didelphis virginiana), and raccoons (Procyon lotor) from Connecticut.

    PubMed

    Mitchell, Sheila M; Richardson, Dennis J; Lindsay, David S

    2006-06-01

    The prevalence of agglutinating antibodies to Toxoplasma gondii was examined in striped skunks (Mephitis mephitis), opossums (Didelphis virginiana), and raccoons (Procyon lotor) from 8 cities in Connecticut. Ten (42%) of the 24 striped skunks, 2 of 7 (29%) opossums, and 12 of 12 (100%) raccoons were positive at dilutions of 1:50 or greater. These results suggest that T. gondii is prevalent in the environment, or prey items, or both, of these omnivores in Connecticut.

  11. Erythrocyte osmotic fragility and oxidative stress in experimental hypothyroidism.

    PubMed

    Dariyerli, Nuran; Toplan, Selmin; Akyolcu, Mehmet Can; Hatemi, Husrev; Yigit, Gunnur

    2004-10-01

    The present study was planned to explain the relation between erythrocyte osmotic fragility and oxidative stress and antioxidant statue in primary hypothyroid-induced experimental rats. Twenty-four Spraque Dawley type female rats were divided into two, as control (n = 12) and experimental (n = 12), groups weighing between 160 and 200 g. The experimental group animals have received tap water methimazole added standard fodder to block the iodine pumps for 30 d (75 mg/100 g). Control group animals were fed tap water and only standard fodder for the same period. At the end of 30 d blood samples were drawn from the abdominal aorta of the rats under ether anesthesia. T3, T4, and TSH levels were measured and the animals that had relatively lower T3, T4, and higher TSH levels were accepted as hypothyroid group. Hormone levels of the control group were at euthyroid conditions. Osmotic fragility, as a lipid peroxidation indicator malondialdehyde (MDA), antioxidant defense system indicators superoxide dismutase (SOD) and glutathione (GSH) levels were measured in the blood samples. Osmotic fragility test results: There was no statistically significant difference found between maximum osmotic hemolysis limit values of both group. Minimum osmotic hemolysis limit value of hypothyroid group was found to be higher than that of control group values (p < 0.02). The standard hemolysis and hemolytic increment curve of the hypothyroid group drawn according to osmotic fragility test results was found to be shifted to the right when compared to control group's curve. This situation and hemolytic increment value, which shows maximum hemolysis ratio, is the proof of increased osmotic fragility of the erythrocytes in hypothyroidism. There is no statistically significant difference found between hypothyroid and control groups in the lipid peroxidation indicator MDA and antioxidant indicators SOD and GSH levels. As a result of our study it may be concluded that hypothyroidism may lead to an

  12. Solubilization of native actin monomers from human erythrocyte membranes.

    PubMed

    Tilley, L; Dwyer, M; Ralston, G B

    1986-01-01

    Up to 50% of the actin in erythrocyte membranes can be solubilized at low ionic strength in a form capable of inhibiting DNAse I, in the presence of 0.4 mM ATP and 0.05 mM calcium. In the absence of calcium and ATP, actin is released but is apparently rapidly denatured. Solubilization of G-actin increases with temperature up to 37 degrees C. At higher temperatures, actin is released rapidly but quickly loses its ability to inhibit DNAse I. PMID:3789986

  13. Malaria parasite pre-erythrocytic stage infection: Gliding and Hiding

    PubMed Central

    Vaughan, Ashley M.; Aly, Ahmed S. I.; Kappe, Stefan H. I.

    2008-01-01

    Summary Malaria is caused by red blood cell-infectious forms of Plasmodium parasites resulting in illness and possible death of infected hosts. The mosquito-borne sporozoite stage of the parasite and the initial infection in the liver, however cause little pathology and no symptoms. Nevertheless, these pre-erythrocytic parasite stages are attracting passionate research efforts not least because they are the most promising targets for malaria vaccine development. Here, we review how the infectious sporozoite makes its way to the liver, subsequently develops in hepatocytes and the factors, both parasite and host, involved in the interactions that occur during this ‘silent’ phase of infection. PMID:18779047

  14. [Rate of erythrocyte sedimentation in buffaloes in related to age].

    PubMed

    d'Angelo, A; Zicarelli, L; Damiano, B; Avallone, L; Crasto, A

    1984-03-30

    The behavior of the ERS was studied in water buffalo of both sexes from one week to nine years of age. A progressive and considerable increase in the values of the ERS, with a respective increase in the animal's age, was noted. Concerning the factors conditioning such peculiarities, the authors attribute importance to the following items: the number of erythrocytes, the plasmatic viscosity, and the total proteinemia (especially in the region of the gamma-globulins that, among those taken in the study, exhibited the greatest variations).

  15. Latex agglutination using the periplasmic proteins antigen of Brucella melitensis is a successful, rapid, and specific serodiagnostic test for ovine brucellosis.

    PubMed

    Ismael, Alaa Bassuny; Swelum, Ayman Abdel-Aziz; Mostafa, Salama A-H; Alhumiany, Abdel-Rahman A

    2016-09-01

    Brucellosis, especially caused by Brucella melitensis, is considered the most-widespread zoonosis in the world, particularly in developing countries. This study was planned to develop an accurate test for diagnosis of ovine brucellosis using a specific hot saline extracted soluble Brucella melitensis periplasmic proteins (SBPPs). The efficacy of the latex agglutination test (LAT) using SBPPs compared to the Rose Bengal test (RBT), buffered plate agglutination test (BPAT), serum agglutination test (SAT), and an indirect enzyme-linked immunosorbent assay (i-ELISA) was evaluated in the field diagnosis of ovine brucellosis. The test performance was evaluated by estimating sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), disease prevalence (DP), positive likelihood ratio (PLR), and negative likelihood ratio (NLR) using test agreement and bacteriological culture in 1777 samples. The false-positive result was significantly (P ⩽0.05) lower in LAT than RBT, BPAT, SAT, and i-ELISA. With reference to test agreement, the Se, Sp, PPV, and PLR were highest (P ⩽0.05) in LAT 99.33%, 99.88%, 98.68%, and 827.25%, respectively. With reference to bacteriological culture, the LAT and i-ELISA tests showed a significant difference in Se with SAT. However, no significant difference in specificity was detected. The DP was 8.44% in the five tests. In conclusion, LAT using SBPPs of B. melitensis could be a suitable serodiagnostic field test for ovine brucellosis, with high sensitivity and specificity. PMID:27207442

  16. Lab-on-a-disc agglutination assay for protein detection by optomagnetic readout and optical imaging using nano- and micro-sized magnetic beads.

    PubMed

    Uddin, Rokon; Burger, Robert; Donolato, Marco; Fock, Jeppe; Creagh, Michael; Hansen, Mikkel Fougt; Boisen, Anja

    2016-11-15

    We present a biosensing platform for the detection of proteins based on agglutination of aptamer coated magnetic nano- or microbeads. The assay, from sample to answer, is integrated on an automated, low-cost microfluidic disc platform. This ensures fast and reliable results due to a minimum of manual steps involved. The detection of the target protein was achieved in two ways: (1) optomagnetic readout using magnetic nanobeads (MNBs); (2) optical imaging using magnetic microbeads (MMBs). The optomagnetic readout of agglutination is based on optical measurement of the dynamics of MNB aggregates whereas the imaging method is based on direct visualization and quantification of the average size of MMB aggregates. By enhancing magnetic particle agglutination via application of strong magnetic field pulses, we obtained identical limits of detection of 25pM with the same sample-to-answer time (15min 30s) using the two differently sized beads for the two detection methods. In both cases a sample volume of only 10µl is required. The demonstrated automation, low sample-to-answer time and portability of both detection instruments as well as integration of the assay on a low-cost disc are important steps for the implementation of these as portable tools in an out-of-lab setting.

  17. Lab-on-a-disc agglutination assay for protein detection by optomagnetic readout and optical imaging using nano- and micro-sized magnetic beads.

    PubMed

    Uddin, Rokon; Burger, Robert; Donolato, Marco; Fock, Jeppe; Creagh, Michael; Hansen, Mikkel Fougt; Boisen, Anja

    2016-11-15

    We present a biosensing platform for the detection of proteins based on agglutination of aptamer coated magnetic nano- or microbeads. The assay, from sample to answer, is integrated on an automated, low-cost microfluidic disc platform. This ensures fast and reliable results due to a minimum of manual steps involved. The detection of the target protein was achieved in two ways: (1) optomagnetic readout using magnetic nanobeads (MNBs); (2) optical imaging using magnetic microbeads (MMBs). The optomagnetic readout of agglutination is based on optical measurement of the dynamics of MNB aggregates whereas the imaging method is based on direct visualization and quantification of the average size of MMB aggregates. By enhancing magnetic particle agglutination via application of strong magnetic field pulses, we obtained identical limits of detection of 25pM with the same sample-to-answer time (15min 30s) using the two differently sized beads for the two detection methods. In both cases a sample volume of only 10µl is required. The demonstrated automation, low sample-to-answer time and portability of both detection instruments as well as integration of the assay on a low-cost disc are important steps for the implementation of these as portable tools in an out-of-lab setting. PMID:27183287

  18. Selective mineral composition, functional test morphology and paleoecology of the agglutinated foraminiferal genus Colominella Popescu, 1998 in the Mediterranean Pliocene (Liguria, Italy)

    NASA Astrophysics Data System (ADS)

    Nicoletta, Mancin; Elena, Basso; Camilla, Pirini; Michael A., Kaminski

    2012-12-01

    Specimens of Colominella (agglutinated Foraminifera) from a Pliocene Mediterranean succession were analysed through a scanning electron microscope (SEM) equipped with an energy-dispersive X-ray spectrometer (EDX) to document their test microstructure. Colominella develops a complex large test with a mostly biserial chamber arrangement, but with the internal chamber lumens partitioned by vertical and horizontal plates that form a labyrinthine structure of alcoves. This internal partition occurs from the first chambers but is completely masked from the outside by the thick wall. The test-wall microstructure is characterized by canaliculi (parapores) that are externally covered by a pavement of agglutinated grains. The mineralogical characterization of the agglutinated grains and the secreted cement shows that the grains are strongly selected as regards to size, arrangement and composition, with the coarse grains placed close to the outer wall. Moreover, these coarse grains, forming a pavement, are made of monocrystalline quartz, whereas the inner part of the skeleton is mostly composed of dolomite. The carbonate cement is less abundant and appears as cloudy light grey areas among the detrital grains. These shell features can be interpreted as functional adaptations to perform kleptoplastidy and/or to house functional photosymbionts, probably induced by stable environmental conditions as in warm shallow waters characterized by low nutrient flux.

  19. Concanavalin A-mediated agglutination and distribution of concanavalin A-binding sites in Acanthamoeba following treatment with colchicine and cytochalasin B.

    PubMed

    Paatero, G; Isomaa, B; Ranninen, T; Wessberg, S

    1983-01-01

    Incubation of Acanthamoeba castellanii (Neff strain) with FITC-ConA (15 micrograms/ml) resulted in the appearance of patches of fluorescence on the amoebae within 2 min of incubation. These patches disappeared following treatment of the amoebae with alpha-MeMan. Pretreatment of the amoebae with colchicine or cytochalasin B or with colchicine and cytochalasin B in combination did not significantly alter the distribution pattern of fluorescence in the amoebae. 2,4-Dinitrophenol and incubation at 4 degrees C on the other hand decreased the degree of patching of the amoebae. Pretreatment with 2,4-dinitrophenol and incubation at 4 degrees C also decreased the ConA-mediated agglutination of the amoebae. No effect on the ConA-mediated agglutination was, however, observed following pretreatment of the amoebae with colchicine and cytochalasin B neither alone nor in combination. Our results indicate that ConA-mediated agglutination and long-range ConA-receptor mobility in the Acanthamoeba are not under the control of structures sensitive to cytochalasin B or colchicine. PMID:6682041

  20. [Spontaneous Cryptosporidium infection in weaned rabbits].

    PubMed

    Pavlásek, I; Lávicka, M; Tůmová, E; Skrivan, M

    1996-12-01

    The first occurrence of Cryptosporidium parvum Tyzzer, 1912 in broiler rabbits in the Czech Republic is reported. The protozoon was determined on the basis of morphometrical parameters of oocysts and of localization of endogenous developmental stages. The dynamics of natural Cryptosporidium infection was studied in a group of 72 young rabbits after weaning (their age ranging from 23-33 to 82-92 days) obtained from six large flocks and used in a feeding experiment. C. parvum was found in rabbits from four farms (Tab. I). Animals under observation were divided into 9 subgroups according to the genotype (Hyla 2000, California White, crosses of New Zealand x California, New Zealand White, Cunistar and Zika) as well as according to the farm of origin. The animals were housed in 28 cages under the conditions of two-floor cage technology. The upper floor consisted of cages housing three head, the lower floor two head each. The animals were fed ad libitum with commercial feed mixture (till the average age of 64.days supplemented with Robenidin as coccidiostat). During the first 10 days of observation pooled samples of droppings from each cage were examinated by flotation-centrifugation method according to Breza (1957) and Pavlásek (1991) in the intervals of three to four days, later one-week intervals. Post mortem scrapings from mucous epithelium taken from young rabbits were examinated (to reveal endogenous developmental stages of C. parvum) together with digesta (to detect oocysts of the protozoon) taken from the full length of the small intestine using method of native preparations and Giemsa stain. In one 37-day dead animal the small and large intestines were examined histologically. The maximum number of young rabbits infected with C. parvum were 30-40 and 33-43 days old (Fig. 1). In animals of this age category the oocysts of the protozoon were found in pooled samples in 11 and 12 cages (39.3 and 42.9%) from totally 28 cages under study. In rabbits of more than 50