scholarly journals Purification and use of limulin: a sialic acid-specific lectin.

1982 ◽  
Vol 30 (9) ◽  
pp. 938-946 ◽  
Author(s):  
V Muresan ◽  
V Iwanij ◽  
Z D Smith ◽  
J D Jamieson
Keyword(s):  
Sialic Acid ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Light And Electron Microscopy ◽  
Sodium Dodecyl ◽  
Limulus Polyphemus ◽  
Pancreatic Acinar Cells ◽  
Renal Tubules ◽  
Neuraminidase Treatment ◽  
Horse Red Blood Cells

A simple and rapid method for the isolation of the sialic acid-specific lectin, Limulus polyphemus hemagglutinin (LPA), from the hemolymph of Limulus polyphemus is described. Declotted hemolymph is adsorbed to an affinity chromatographic column consisting of hog gastric mucin glycopeptides coupled to agarose and LPA is eluted in a single step with a Ca2+-free buffer, giving an apparent purification of approximately 25,000-fold. The eluted material is homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing and consists of identical subunits each of 29,000 daltons. Hemagglutination inhibition studies with horse red blood cells indicate specificity of LPA for N-acetyl- and N-glycolylneuraminic acid; binding is Ca2+ -dependent and abolished by neuraminidase treatment. LPA was covalently coupled to rhodamine and to horseradish peroxidase for use in detection of sialoglycoconjugates on cells and tissues by light and electron microscopy. Examples of the use of LPA for detection of sialoglycoconjugates in rat renal tubules and glomeruli, blood vessels in rat pancreas, and on horse red blood cells are shown. The procedures described here should prove useful as a cytochemical probe for detection of sialoglycoconjugates in a variety of systems. An accompanying article utilizes these probes for the detection of sialoglycoconjugates on the plasmalemma of adult and differentiating rat pancreatic acinar cells.

scholarly journals Effects of pronase and neuraminidase treatment on a myelin-associated glycoprotein in developing brain

1976 ◽  
Vol 156 (1) ◽  
pp. 143-150 ◽  
Author(s):  
R H Quarles
Keyword(s):  
Molecular Weight ◽  
Sialic Acid ◽  
Sodium Dodecyl Sulphate ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Weight Class ◽  
Developmental Difference ◽  
Adult Rats ◽  
Neuraminidase Treatment

Rats (14 days old) were injected with [14c]fucose and young adult rats with [3H]fucose in order to label the myelin-associated glycoproteins. As previously reported, the major [14C]fucose-labelled glycoprotein in the immature myelin had a higher apparent molecular weight on sodium dodecyl sulphate/polyacrylamide gels that the [3H]fucose-labelled glycoprotein in mature myelin. This predominant doubly labelled glycoprotein component was partially purified by preparative gel electrophoresis and converted to glycopeptides by extensive Pronase digestion. Gel filtration on Sephadex G-50 separated the glycopeptides into several clases, which were designted A,B, C AND D, from high to low molecular weight. The 14C-labelled glycopeptides from immature myeline were enriched in the highest-molecular-weight class A relative to the 3H-labelled glycopeptides from mature myelin. Neuraminidase treatment of the glycoprotein before Pronase digestion greatly decreased the proportion of glycopeptides fractionating in the higher-molecular-weight classes and largely eliminated the developmental differences that were apparent by gel filtration. However, neuraminidase treatment did not decrease the magnitude of the developmental difference revealed by electrophoresing the intact glycoprotein on sodium dodecyl sulphate gels, although it did decrease the apparent molecular weight of the glycoprotein from both the 15-day-old and adult rats by an amount comparable in magnitude to that developmental difference. The results from gel filtration of glycopeptides indicate that there is a higher content of large molecular weight, sialic acid-rich oligosaccharide units in the glycoprotein of immature myelin. However, the higher apparent molecular weight for the glycoprotein from 15-day-old rats on sodium dodcyl sulphate gels is not due primarily to its higher sialic acid content.

scholarly journals The Effect of Experimental Escherichia coli Infection on Some Blood Parameters and Histological Changes in Male Rats

2011 ◽  
Vol 35 (2) ◽  
pp. 22-27
Author(s):  
Hayder AL – Zamely
Keyword(s):  
Escherichia Coli ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Histopathological Examination ◽  
Hemoglobin Concentration ◽  
Blood Parameters ◽  
Infected Group ◽  
Male Rats ◽  
Control Group ◽  
Renal Tubules

This study was carried out to investigate the effect of experimental infection by Escherichia coli on some blood parameters and histopathological changes in male rats to reach this objective twenty male rats (Rattus norvigicus) were divided into two groups, housed under same conditions of water, feed, temperature, light and humidity. The first one served as control group, while the second was experimentally infected with Ecoli by administration of 2ml / rat of (1×109) colony forming unit/ml of E-coli. Five days later blood were obtained from animals by cardiac puncture to measure the total and differential white blood cells count hemoglobin concentration packed cells volume red blood cells count, and thrombocytes count. One animal from each group was killed and histopathological examination was done for intestine kidney liver and spleen to detect the effect of infection on these organs. The results revealed a significant increase in total leukocytes count (P<005) in infected group while the differential count showed a significant increase in neutrophils and significant decrease in lymphocytes (P< 0.05) in same group. There were a significant decrease in hemoglobin concentration packed cell volume and non significant decrease in red blood cells count. While thrombocytes count showed a significant decrease in infected group compared with control group (P<0.05). The histopathological examination of infected male rats intestine was demonstrated mild diffuse inflammation which include increased numbers of neutrophile in tissue intraepithelial lymphocytes and goblet cells In kidney renal tubules showed extensive epithelial swelling with decreased lumen space and generalized necrotic changes with interstitial hemorrhage in renal cortex. In liver hepatocytes were swollen with decreased sinusoidal spaces and widely distributed necrotic foci were seen. There is extensive splenic necrosis with infiltration of leukocytes from white pulp to red pulp of spleen.

scholarly journals Spherization of red blood cells and platelets margination in COPD patients

2020 ◽  
Author(s):  
Karim Zouaoui Boudjeltia ◽  
Christos Kotsalos ◽  
Daniel Ribeiro ◽  
Alexandre Rousseau ◽  
Christophe Lelubre ◽  
...  
Keyword(s):  
Shear Rate ◽  
Numerical Simulations ◽  
Red Blood Cells ◽  
Conformational Changes ◽  
Blood Cells ◽  
Mean Square Displacement ◽  
Neuraminidase Treatment ◽  
Copd Patients ◽  
Platelet Aggregates

AbstractRationaleThere are important interactions between Red Blood Cells (RBCs) and platelets in the bloodstream. These interactions lead to a phenomenon called margination. RBCs in pathological situations undergo biochemical and conformational changes leading to alterations in blood rheology.AimRBCs shape in volunteers (21), stable (42) and exacerbated (31) COPD patients was analyzed. We studied the effect of the RBCs spherization on the platelets transport experimentally, in vitro, and by using numerical simulations.MethodsRBC shape was estimated by the second moment of Pearson obtained through flow cytometry on fsc histogram. In vitro experiments were performed to analyze the effect of RBC shape on platelets adhesion/aggregation in dynamic conditions. Neuraminidase treatment was used to induce RBCs spherization. Numerical simulation were performed to determine the effect of RBCs spherization on platelets mean square displacement (MSD) to provide a physical explanation.ResultsSignificant increase of RBC sphericity was observed in COPD patients compared to volunteers (Kruskal-Wallis: p<0.0001). In vitro experiments, at shear rate of 100 s-1, we observed that RBCs treated with neuraminidase mainly affect the number of platelet aggregates (p = 0.004). There was no change in the aggregates size. At a shear rate of 400 sec-1 neuraminidase treatment changes both the size of the aggregates (p = 0.009) and the number of platelet aggregates (p = 0.008).Numerical simulations indicated that RBCs spherization induces an increase of MSD and the effect was more pronounced when the shear rate increased.ConclusionOur results show that the RBCs of COPD patients are more spherical than those of healthy volunteers. Experimentally we observe that the RBCs spherization induces an increase platelet transport to the wall. Additional studies are needed to better understand the possible association between the RBCs effect on the platelets transport and the increased cardiovascular events observed in COPD patients.

scholarly journals Restoring the youth of aged red blood cells and extending their lifespan in circulation by remodelling membrane sialic acid

2016 ◽  
Vol 20 (2) ◽  
pp. 294-301 ◽  
Author(s):  
Yao‐Xiong Huang ◽  
Wei‐Wei Tuo ◽  
Di Wang ◽  
Li‐Li Kang ◽  
Xing‐Yao Chen ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Red Blood Cells ◽  
Blood Cells

scholarly journals Expression of sialic acid on the alveolar surface of adult and fetal human lungs.

1986 ◽  
Vol 34 (6) ◽  
pp. 811-816 ◽  
Author(s):  
T Faraggiana ◽  
D Villari ◽  
J Jagirdar ◽  
J Patil
Keyword(s):  
Sialic Acid ◽  
Light And Electron Microscopy ◽  
Fetal Lung ◽  
Peanut Lectin ◽  
Neuraminidase Treatment ◽  
Human Lungs ◽  
Type Ii Pneumocytes ◽  
Respiratory Tree ◽  
Binding Lectin ◽  
Alveolar Surface

Lung alveoli are coated by a thin layer of extracellular material rich in anionic charges. The nature of this acid layer and its relationship to the phospholipid surfactant are not known. We investigated the possible presence of sialic acid groups by light and electron microscopy in tissues from normal fetal and adult lungs, using neuraminidase treatment followed by staining with the galactose-binding lectin from peanut, labeled with peroxidase. Our results showed that adult lung does not bear peanut lectin-reactive sites but that a very thin and distinct reactive layer becomes evident after neuraminidase treatment, especially on type II pneumocytes. In fetal lung, the entire surface of the developing respiratory tree is outlined by a strongly peanut lectin-reactive layer even if neuraminidase digestion is not performed. We conclude that the acid coat of the alveolar lining is in part composed of sialic acid residues and that sialic acid is added to the fetal lung as the alveoli mature.

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