PubMed Health⌕ Search

Biomedical subjects

G S Chhatwal

Publications and source records attributed to G S Chhatwal.

86 records · Page 5Linked to original sources
PubMed1984

Guanidine extraction enhances the binding of human fibrinogen to group-B streptococci.

Treatment of group-B streptococci with guanidine chloride significantly enhanced their capacity to bind 125I-labelled fibrinogen. The increase in binding activity was almost proportional in the range of 0.05 M to 6 M guanidine chloride. Repeated extractions with guanidine chloride further increased the capacity of B streptococci to bind the labelled fibrinogen. On the other hand, urea, even at concentrations up to 8 M, did not alter the fibrinogen binding. The enhancement was most pronounced with B streptococci of serotypes III and III-R. Cultivation of these streptococci in Baker and Kasper medium, which is known to stimulate formation of microcapsules, reduced 125I-fibrinogen binding. Subsequent treatment with guanidine increased their binding activity, presumably by facilitating the accessibility of the binding sites.

Binding Sites↗
PubMed1984

Characterization of immunoglobulin G binding to Staphylococcus aureus strain Wood 46.

Protein A (PA)-activity was detected in Staphylococcus aureus strain Wood 46 which had been considered to be PA-negative. This staphylococcal strain bound 28% of 125I-labelled IgG, compared with 89% by strain Cowan I. The binding activities of both S. aureus strains were saturable, time-dependent and specific. The dissociation constants of 1.6 X 10(-9) M for Wood 46 and 9.3 X 10(-8) M for Cowan I indicated a similar affinity for human IgG in both strains. The number of IgG-binding sites were estimated to be 16,970 for Wood 46 and 41,200 for Cowan I. Exposure to heat and ultrasonication reduced PA-activities of strain Cowan I, but not that of strain Wood 46. Extraction of the staphylococci with guanidine and formic acid resulted in a reduction of IgG-binding activities only in strain Wood 46. Photooxidation, trypsinization and lysozyme treatment also diminished IgG-binding of strain Wood 46 to a larger extent than that of strain Cowan I. Extracellular PA from S. aureus strains Wood 46 and Cowan I could be purified by affinity chromatography on IgG-sepharose. The purified PA preparations gave single protein bands upon SDS-polyacrylamide gel electrophoresis. Their molecular weights were 42,000 and their isoelectric points approximated 5.0.

Binding Sites, Antibody↗
PubMed1983

Binding of human fibrinogen and its polypeptide chains to group B streptococci.

Of 51 group-B streptococcal cultures, 20 bound 125I-labelled human fibrinogen. In contrast to the streptococci of groups A, C and G, the group-B streptococci interacted more with the alpha, beta-chain of fibrinogen than with whole fibrinogen. None of the streptococcal cultures reacted with gamma-chain. The fibrinogen-negative group-B streptococci still bound the alpha, beta-chain. The binding of 125I-labelled, alpha, beta-chain could be inhibited by unlabelled fibrinogen with fibrinogen-positive, but not with fibrinogen-negative streptococci of group B.

Alkylation↗
PubMed1983

Variations in the binding of mammalian fibrinogens to streptococci of different animal origin.

Binding of 125I-labelled fibrinogen from humans, bovines, equines, canines and ovines by streptococci of serological groups A, B, C and G was determined quantitatively. All 59 randomly selected streptococcal cultures generally bound more human fibrinogen than the other fibrinogens. Only Sc. dysgalactiae had a higher affinity for bovine fibrinogen. In addition, Sc. dysgalactiae bound distinctly more equine, canine and ovine fibrinogen than the other streptococci. Some cultures of Sc. equi and Sc. zooepidemicus had high binding activities for equine fibrinogen. Low binding capacities were exhibited by B-streptococci, particularly with fibrinogens from horses and dogs.

Animals↗
PubMed1983

Characterization of binding of human alpha 2-macroglobulin to group G streptococci.

An interaction was observed between human alpha 2-macroglobulin (alpha 2M) and streptococci belonging to group A, C, and G. Of 27 group C and 19 group G streptococcal cultures, 13 and 14, respectively, bound 125I-labeled alpha 2M. Some group A streptococci also interacted with alpha 2M. A number of other bacterial species tested did not react with alpha 2M. The binding of 125I-labeled alpha 2M to group G streptococci was time dependent, saturable, and could be inhibited by unlabeled alpha 2M. Inhibition experiments indicated that the streptococcal binding site for alpha 2M differed from the receptors for immunoglobulin G, fibrinogen, aggregated beta 2-microglobulin, albumin, and fibronectin. The alpha 2M binding activity was remarkably sensitive to trypsin and heat treatment indicating its protein nature. Kinetic analysis indicated a homogenous population of binding sites. The number of binding sites per bacterial cell was estimated to be approximately 20,000.

Binding Sites↗
PubMed1982

Changes in erythrocyte permeability due to palytoxin as compared to amphotericin B.

Palytoxin causes within minutes a temperature-dependent K+ loss from human and rat erythrocytes which is followed within hours by haemolysis. It decreases the osmotic resistance in a concentration-dependent manner, so that osmotic influences are negligible for K+ release but considerable in haemolysis. External K+ inhibits the haemoglobin release and Rb+ inhibits the release of K+ and haemoglobin. Ca2+ (over 20 microM) and borate (over 5 microM) enhance the loss of K+ and haemoglobin. With both Ca2+ and borate present, the efficacy of palytoxin is raised about 10 000-fold. Under these conditions, about 15 palytoxin molecules per human cell trigger a 50% K+ loss over a wide range of cell concentrations. The palytoxin effect is reversible. After depletion from K+ by low concentrations of palytoxin, human cells can be refilled with K+ and resealed. The pores formed by palytoxin are small. They allow the entrance of Na+ and choline, whereas inositol is largely excluded and Ca2+, as well as sucrose and inulin, are completely excluded. Amphotericin B resembles palytoxin in its ability to cause a considerable prelytic K+ loss and to form small pores. However, it is about 1000-times weaker than palytoxin, is not inhibited by K+ or Rb+, is not activated by Ca2+ or borate, and has a negative temperature dependence. Thus palytoxin represents a novel type of cytolysin.

Acrylamides↗
PubMed1982

Ouabain inhibits the increase due to palytoxin of cation permeability of erythrocytes.

1. Palytoxin in concentrations as low as 1 pM raises the potassium permeability of rat, human and sheep erythrocytes, and the sodium permeability of human erythrocytes. The release of potassium or sodium from human cells also occurs when extracellular sodium is replaced by choline. 2. Ouabain inhibits the release due to palytoxin of potassium ions from human, sheep and rat erythrocytes, and also the release of sodium ions from human cells. The glycoside effect is specific since a) it is already prominent with 5 X 10(-8) M ouabain b) rat erythrocytes are less sensitive than human cells to ouabain c) potassium release due to amphotericin B or the Ca2+ ionophore A 23187 is not influenced by ouabain and d) dog erythrocytes are resistant to palytoxin as well as to ouabain. 3. Palytoxin has no direct influence on the Na+, K+ - ATPase. It inhibits the binding of [3H]ouabain to erythrocyte membranes within the same concentration range as unlabelled ouabain. It partially displaces bound [3H]ouabain, and partially inhibits the inactivation of erythrocyte ATPase by the glycoside. Depletion of ATP or of external Ca2+ renders the cells less sensitive to palytoxin. Nevertheless inhibition by ouabain can be still demonstrated with human cells whose ATP stores had been largely exhausted, and also in the absence of external Ca2+. 4. Palytoxin decreases the surface tension at the air-water interface. We assume that the formation of nonspecific pores by palytoxin is linked with its surface activity. Further experiments should demonstrate whether ouabain prevents the binding of palytoxin to erythrocytes ("receptor hypothesis"), or whether an ouabain-sensitive hydrolysis of trace amounts of ATP ("metabolic hypothesis") promotes the palytoxin effect.

Acrylamides↗
PubMed1982

Palytoxin both induces and inhibits the release of histamine from rat mast cells.

Palytoxin (PTX) is a potent releaser of histamine from rat mast cells. The concentration response curve is bell-shaped with its range between 0.05 ng/ml and 5 micrograms/ml and its maximum at 50 ng/ml. The release of histamine by PTX is specific because (a) heat-inactivated (50 degrees C) mast cells are insensitive to PTX, and (b) extracellular calcium is essential. Removal of extracellular potassium as well as the presence of borate shifts the curve to the left by factor of 10. At higher concentrations (5 micrograms/ml), PTX inhibits the release of histamine due to compound 48/80, MCD-peptide, Con-A, ionophore A-23187, and PTX itself.

Acrylamides↗
PubMed1981

Delayed haemolytic action of palytoxin. General characteristics.

1. Palytoxin is a haemolysin. The erythrocytes from various species can be classified into a sensitive and a hardly sensitive group. The former contain potassium as their main inside cation and are arranged according to their sensitivity as hog greater than or equal to rat, mouse greater than rabbit greater than guinea-pig greater than man. The latter, comprising those from sheep and cattle, have sodium as their main inside cation. In addition, chicken erythrocytes are relatively insensitive. 2. Haemolysis of rat erythrocytes is preceded by a lag period of 1--2 h. With increasing temperature the haemolysis proceeds more quickly but reaches the same final range between 25 and 42 degrees C. The pH optimum in Britton-Robinson buffer supplemented with saline is between 7 and 8. Washing off palytoxin during the prelytic period reduces the haemolytic power. 3. The sensitivity of rat erythrocytes decreases with increase of osmolarity between 235 and 415 mosM. Accordingly, their osmotic resistance is lowered by palytoxin in a concentration-dependent manner. 4. With both rat and sheep erythrocytes, potassium loss by far precedes the haemolysis due to palytoxin. Potassium loss is measurable already after 1 min and increases with time. After 2 hours the quotient between the ED50 of haemolysis and that of potassium loss is around 200. Thus palytoxin is an unusually strong but slow haemolysin of the osmotic type. The extreme prelytic potassium loss and the correlation between susceptibility and potassium content of erythrocytes points towards the relevance of ionic fluxes.

Acrylamides↗
PubMed1978

Purification of a nontoxic phospholipase A2 from the venom of Indian krait (Bungarus caeruleus).

A nontoxic phospholipase A2 was purified from the venom of Indian krait (Bungarus caeruleus) by a four-step procedure involving electrophoresis, gel filtration and ion-exchange chromatography. The recovery of the enzyme activity was 37% and the purified preparation was 38 times as active as the crude venom. The purified enzyme had a molecular weight of 12,500 and the optimum pH of 7.2. The enzyme showed higher specificity toward phosphatidylethanolamine than phosphatidylcholine. The preparation was not very labile to heat and its activity was dependent on the presence of divalent cations, calcium ions being the most effective activators. The enzyme was completely inhibited by iodoacetic acid but showed high stability against 8 M urea. Purified phospholipase A2 was nontoxic at an iv dose of 5 microgram/g mouse. The high specific activity, the high yield and the nontoxic nature of the enzyme indicate that the major form of phospholipase A2 in Bungarus caeruleus venom is not associated with any toxicity and has properties somewhat similar to that of phospholipase A2 from some other venoms.

Animals↗