scholarly journals THE CRYSTALLIZATION AND SEROLOGICAL DIFFERENTIATION OF A STREPTOCOCCAL PROTEINASE AND ITS PRECURSOR

1950 ◽  
Vol 92 (3) ◽  
pp. 201-218 ◽  
Author(s):  
S. D. Elliott
Keyword(s):  
Ammonium Sulfate ◽  
Crystalline Form ◽  
Antigenic Specificity ◽  
Serological Tests ◽  
Saturated Ammonium Sulfate ◽  
Alkaline Reaction ◽  
Group A ◽  
Refrigerator Temperature ◽  
Streptococcal Proteinase ◽  
Degree Of Purification

Grown in dialysate broth at a pH between 5.5 and 6.5, some strains of group A streptococci elaborate the precursor of a proteolytic enzyme. Within this range of hydrogen concentration the precursor is also produced when the streptococci are suspended in a peptone dialysate containing glucose and incubated at 37°C. The precursor does not appear to be produced at a neutral or alkaline reaction. Methods are described whereby the precursor and proteinase have been isolated in crystalline form. The precursor crystallizes from half-saturated ammonium sulfate at pH 8.0 and a temperature of 22°C. or higher; the proteinase crystallizes from 0.15 saturated ammonium sulfate at pH 8.0 but does so most readily at refrigerator temperature. The degree of purification achieved by these procedures is discussed. The activity of purified preparations of the precursor and of proteinase has been tested against α-benzoyl-l-arginineamide and, with this as a substrate, the conversion of precursor to proteinase by autocatalysis or by trypsin has been confirmed. Immunological experiments are described, the results of which provide evidence of the distinct antigenic specificity of the precursor and proteinase; the conversion of precursor to proteinase has been followed by means of serological tests.

MECHANISMS OF PLATELET AGGREGATION BY ACIDIC MUCOPOLYSACCHARIDE EXTRACTED FROM STICHOPUS JAPONICUS SELENKA IN HUMANS AND RABBITS

1987 ◽  
Author(s):  
J Z Li ◽  
E C-Y Lian
Keyword(s):  
Platelet Aggregation ◽  
Ammonium Sulfate ◽  
Human Platelets ◽  
Platelet Inhibitors ◽  
Rabbit Plasma ◽  
Human Fibrinogen ◽  
Stichopus Japonicus ◽  
Saturated Ammonium Sulfate ◽  
Rabbit Platelets ◽  
Induced Aggregation

It has been reported that acidic mucopolysaccharide extracted from sea cucumber (Stichopus japonicus selenka) (SJAMP) induced the aggregation of human and animal platelets by an unknown mechanism, using platelet-rich plasma (prp) and washed human and rabbit platelets we studied the effects of storage, platelet inhibitors, and various plasmas and their fractions on SJAMP-induced platelet aggregation. we found that the lowest concentrations of SJAMP required for aggregation of human and rabbit platelets were 0.4 and 2 ug/ml respectively. The reactivity of human platelets to SJAMP decreased with time after drawing of blood; rabbit platelets did not show this phenomenon. Platelet inhibitors such as aspirin, indomethacin, apyrase, antimycin, 2-deoxy-D-glucose, and EDTA inhibited by 50 to 100% the aggregation of human platelets induced by SJAMP; but these inhibitors had no effect on SJAMP-induced aggregation of rabbit platelets. Washed human and rabbit platelets were not aggregated by SJAMP. The aggregation of washed human platelets by SJAMP was restored completely by human or rabbit plasma, by human fibrinogen, or by 0 to 30% saturated ammonium sulfate fraction but not by serum. The aggregation of rabbit platelets by SJAMP could only be restored by rabbit plasma or serum, or by 50 to 60% saturated ammonium sulfate fraction. The data indicate that the mechanisms of aggregation of human and rabbit platelets by SJAMP are different. THe SJAMP-induced human platelet aggregation is dependent upon metabolism, release of ADP and the cyclooxygenase pathway requiring fibrinogen and Ca++. The aggregation of rabbit platelets induced by SJAMP is independent of metabolism, release of ADP and cyclooxygenase pathway, and does not require fibrinogen and Ca++, but needs certain protein(s) in the 50 to 60% saturated ammonium sulfate fraction of rabbit plasma.

Intrafascicular Injection of Ammonium Sulfate and Bupivacaine in Peripheral Nerves of Neonatal and Juvenile Rats

1998 ◽  
Vol 23 (2) ◽  
pp. 152-158 ◽  
Author(s):  
M. Catherine Hertl ◽  
Patricia K. Hagberg ◽  
Daniel A. Hunter ◽  
Susan E. Mackinnon ◽  
Jacob C. Langer
Keyword(s):  
Ammonium Sulfate ◽  
Peripheral Nerves ◽  
Animal Studies ◽  
Age Groups ◽  
Age Group ◽  
Nerve Blocks ◽  
Provide Pain Relief ◽  
Group A ◽  
Old Rats ◽  
Track Analysis

Background and ObjectivesRegional nerve blocks are often used for the treatment of postoperative pain in children. Ammonium sulfate is a non-narcotic anesthetic agent, which has been reported to provide pain relief lasting days to weeks, with few reported side effects in adult studies. Prior to considering clinical use in children, the neurotoxicity of ammonium sulfate in 4-day and 3-week old rats was assessed and compared with that of bupivacaine.MethodsEach rat received a posterior tibial nerve intrafascicular injection (0.01 mL in 4-day-old and 0.02 mL in 3-week-old rats) using either 10% ammonium sulfate (n = 24 per age group), 0.5% bupivacaine (n = 18 per age group), 0.9% saline (n = 18 per age group), or 5% phenol (n = 18 per age group). A functional assessment by serial walking track analysis and a morphologic assessment by neurohistology were made.ResultsNo abnormalities in serial walking track analysis and no structural nerve damage were detected after ammonium sulfate, bupivacaine, or saline injection. Bupivacaine caused mild focal changes in both age groups, which recovered by 8 weeks.ConclusionsIntrafascicular injection of ammonium sulfate was as safe as bupivacaine in this animal model. Further animal studies must be made before human trials are initiated.

scholarly journals BACTERICIDAL SUBSTANCE FROM STAPHYLOCOCCUS AUREUS

1970 ◽  
Vol 131 (5) ◽  
pp. 1004-1015 ◽  
Author(s):  
Adnan S. Dajani ◽  
Ernest D. Gray ◽  
Lewis W. Wannamaker
Keyword(s):  
Staphylococcus Aureus ◽  
Ammonium Sulfate ◽  
Phage Type ◽  
Gel Filtration ◽  
Considerable Degree ◽  
Ammonium Sulfate Fractionation ◽  
Sulfate Fractionation ◽  
Extracellular Products ◽  
Group Ii ◽  
Degree Of Purification

A bactericidal substance previously isolated from phage type 71 Slaphylococcus aureus has been further identified and characterized. Staphylococci belonging to phage type 71 produce the substance in higher titers than staphylococci lysed by other phages in group II in addition to phage 71. Other staphylococci do not produce the bactericidal substance. The bactericidal substance shares several of the properties of bacteriocins but differs from this group of antibiotic substances in some respects. A combination of ammonium sulfate fractionation and gel filtration on a Sephadex G-100 column resulted in considerable degree of purification of the bactericidal substance. The substance is a previously unrecognized product of S. aureus and is distinct from other extracellular products of this organism.

Crambe Thioglucoside Glucohydrolase (EC 3.2.3.1): Separation of a Protein Required for Epithiobutane Formation

1973 ◽  
Vol 51 (12) ◽  
pp. 1654-1660 ◽  
Author(s):  
H. L. Tookey
Keyword(s):  
Ammonium Sulfate ◽  
Ferrous Ion ◽  
Seed Meal ◽  
Proteinaceous Material ◽  
Saturated Ammonium Sulfate ◽  
Crambe Seed

Sonicated extract of crambe seed meal prepared in the presence of ferrous ion and dithiothreitol enzymatically converts epi-progoitrin to glucose, HSO4−, and a mixture of 1-cyano-2-hydroxy-3,4-epithiobutanes (50–70%) and 1-cyano-2-hydroxy-3-butene (30–50%). A fraction of the extract precipitating between 60 and 70% saturated ammonium sulfate contains thioglucosidase that converts epi-progoitrin essentially to 1-cyano-2-hydroxy-3-butene. Chromatography (on cross-linked dextran) of a 40–60% ammonium sulfate fraction leads to separation of a proteinaceous material (s20 = 2.6 S) that does not hydrolyze epi-progoitrin but, in the presence of thioglucosidase, promotes the formation of 1-cyano-2-hydroxy-3,4-epithiobutanes in amounts proportional to those from crude seed meal extract.

scholarly journals VARIATION OCCURRING IN GROUP A STREPTOCOCCI DURING HUMAN INFECTION

1948 ◽  
Vol 87 (6) ◽  
pp. 521-533 ◽  
Author(s):  
Sidney Rothbard ◽  
Robert F. Watson
Keyword(s):  
T Antigen ◽  
Human Infection ◽  
M Protein ◽  
Group A Streptococci ◽  
Upper Respiratory Tract ◽  
Group A ◽  
Normal Human ◽  
Streptococcal Proteinase ◽  
Relationship Of ◽  
Observation Period

A study was made of the variation occurring in group A streptococci during the natural course of infection in man. From 54 patients with 56 different group A streptococcal infections of the upper respiratory tract, 251 strains of streptococci, isolated at weekly intervals following infection, were tested for their capacity to resist the bacteriostatic action of normal human blood. In 52 of the infections the streptococci were of recognized serological types and were also tested for variation in their ability to produce the type-specific M protein antigen. Strains isolated in the 1st week of infection were uniformly highly resistant to bacteriostasis and elaborated large amounts of M substance. In 42 per cent of the 52 infections, strains isolated in the convalescent and carrier stages showed an increasing susceptibility to bacteriostasis correlated with a progressive loss of M substance; whereas in the remaining 58 per cent resistance to bacteriostasis and the capacity to produce M protein were maintained throughout the observation period. In 3 different infections, the streptococci became so degraded that no M protein could be demonstrated in acid extracts of these variants. Concomitantly these strains became highly susceptible to bacteriostasis. Spontaneous reversion did not occur, but serial mouse passage reestablished these functions. These degraded variants had the same T antigen as their respective original strains. No evidence was obtained that variation of group A streptococci in resistance to bacteriostasis or in the ability to produce the type-specific M antigen was associated (a) with the appearance of type-specific bacteriostatic antibodies; (b) with any particular serological type of streptococcus; (c) with the production of streptococcal proteinase which digests the M protein; (d) with the therapeutic administration of sulfadiazine; or (e) with the development of complications. The possible relationship of these observations to the problem of the "dangerous carrier" of group A hemolytic streptococci is discussed.

scholarly journals CRYSTALLINE PNEUMOCOCCUS ANTIBODY

1949 ◽  
Vol 32 (6) ◽  
pp. 705-724 ◽  
Author(s):  
John H. Northrop ◽  
Walther F. Goebel
Keyword(s):  
Ammonium Sulfate ◽  
Horse Serum ◽  
Insoluble Fraction ◽  
Rabbit Serum ◽  
Type I ◽  
Type Ii ◽  
Serum Globulin ◽  
Saturated Ammonium Sulfate ◽  
Diphtheria Antitoxin ◽  
Specific Polysaccharide

1. The immune precipitate formed by antipneumococcus horse serum and the specific polysaccharide is not hydrolyzed by trypsin as is the diphtheria toxin-antitoxin complex, and purified pneumococcus antibody cannot be isolated by the method used for the isolation and crystallization of diphtheria antitoxin. 2. Type I pneumococcus antibody, completely precipitable by Type I polysaccharide, may be obtained from immune horse serum globulin by precipitation of the inert proteins with acid potassium phthalate. 3. The antibody obtained in this way may be fractionated by precipitation with ammonium sulfate into three main parts. One is insoluble in neutral salts but soluble from pH 4.5 to 3.0 and from pH 9.5 to 10.5. This is the largest fraction. A second fraction is soluble in 0.05 to 0.2 saturated ammonium sulfate and the third fraction is soluble in 0.2 saturated ammonium sulfate and precipitated by 0.35 saturated ammonium sulfate. The second fraction can be further separated by precipitation with 0.17 saturated ammonium sulfate to yield a small amount of protein which is soluble in 0.17 saturated ammonium sulfate but insoluble in 0.25 saturated ammonium sulfate. This fraction crystallizes in poorly formed, rounded rosettes. 4. The crystallization does not improve the purity of the antibody and is accompanied by the formation of an insoluble protein as in the case of diphtheria antitoxin. 5. None of the fractions obtained is even approximately homogeneous as determined by solubility measurements. 6. Purified antibody has also been obtained by dissociating the antigen-antibody complex. 7. The protective value of the fractions is quite different; that of the dissociated antibody being the highest and that of the insoluble fraction, the lowest. 8. All the fractions are immunologically specific since they do not precipitate with Type II polysaccharide nor protect against Type II pneumococci. 9. All the fractions give a positive precipitin reaction with antihorse rabbit serum. The dissociated antibody gives the least reaction. 10. Comparison of the various fractions, either by their solubility in salt solution or through immunological reactions, indicates that there are a large number of proteins present in immune horse serum, all of which precipitate with the specific polysaccharide but which have very different protective values, different reactions with antihorse rabbit serum, and different solubility in salt solutions.

scholarly journals Safety and Humoral Responses to SARS-CoV-2 mRNA Vaccination of Previously Infected and Naive Populations

2021 ◽  
Author(s):  
Shai Efrati ◽  
Merav Catalogna ◽  
Ramzia Abu Hamed ◽  
Amir Hadanny ◽  
Adina Bar-Chaim ◽  
...  
Keyword(s):  
Single Dose ◽  
Vaccine Dose ◽  
Medical Attention ◽  
Evaluation Procedure ◽  
Short Term ◽  
Serological Tests ◽  
Safety Issues ◽  
Infected Population ◽  
Group A ◽  
Humoral Responses

Abstract Since COVID-19 risk of reinfection is of great concern, the safety and efficacy of the mRNA-based vaccines in previously infected populations should be assessed. We studied 78 individuals previously infected with SARS-CoV-19, who received a single dose of BNT162b2 mRNA COVID-19 vaccine, and 1:2 ratio matched infection-naïve cohort who received two injections. The evaluation procedure included symptom monitoring, and serological tests. Among the post-infected population, the median IgG-S response after the first vaccine dose was 2260 AU/ml, compared to 238 AU/ml after the second vaccine injection in the infection naive group. A strong correlation was demonstrated between IgG-S level before vaccination, and the corresponding responses after a single vaccine dose (r = 0.8, p < 0.001) in the post infected population. Short-term severe symptoms that required medical attention were found in 6.8% among the post-infected individuals, while none were found in the infection naïve population. Our data suggest that a single vaccine dose is sufficient to induce an intense immune response in post-infected population regardless of seropositivity. Although some short-term safety issues were observed compared to the infection naïve population, a single dose regimen can be considered safe in post-infected populations.

GROUP A STREPTOCOCCAL INFECTIONS IN A CHILDREN'S HOME

PEDIATRICS ◽  
1964 ◽  
Vol 33 (1) ◽  
pp. 5-10
Author(s):  
Hugh L. Moffet ◽  
Henry G. Cramblett ◽  
Joyce P. Black
Keyword(s):  
Agar Plate ◽  
Streptococcal Pharyngitis ◽  
School Age Children ◽  
Blood Agar Plate ◽  
Normal Children ◽  
Simple Method ◽  
Sheep Blood Agar Plate ◽  
Children's Home ◽  
Group A ◽  
Refrigerator Temperature

School-age children living in a children's home had pharyngeal cultures made on 485 consecutive infirmary admissions during a period of 14 months by inoculating the throat swab on the surface of a sheep blood agar plate. ASO titers were determined for the acute and convalescent phases of 95% of the 321 untreated illnesses associated with negative cultures. A rise in ASO titer occurred in 3.6% of these illnesses. ASO titers were also determined on pairs of sera for 51 of 53 untreated febrile illnesses associated with negative streptococcal cultures in 1962 and a rise in ASO titer occurred for only one of these illnesses (2%). This simple method for pharyngeal cultures is adequate and accurate as a laboratory aid to the practicing physician for the diagnosis of streptococcal pharyngitis in normal children. Bacitracin sensitivity showed an excellent correlation with Group A streptococci. Studies of duplicate swabs indicated that overnight storage of a dry swab at room or refrigerator temperature was associated with the recovery of more than 10 colonies of hemolytic streptococci from 80% to 88% of the swabs whose duplicates had more than 50 colonies.

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