scholarly journals A Hypertransfused Mouse Assay for Thrombopoietic Factors

Blood ◽  
1973 ◽  
Vol 42 (3) ◽  
pp. 423-428 ◽  
Author(s):  
George W. Cooper ◽  
Barbara Cooper ◽  
A. Lawrence Ossias ◽  
Esmail D. Zanjani
Keyword(s):  
Human Serum ◽  
Whole Blood ◽  
Hodgkin's Disease ◽  
Healthy Donor ◽  
Hodgkin’S Disease ◽  
Normal Serum ◽  
Normal Platelet ◽  
Quantitative Separation ◽  
Thrombocytopenic Patient ◽  
Normal Human

Abstract A method was developed for the quantitative separation of platelets from CF1 mouse whole blood. This made it possible to determine the platelet incorporation of 35S-sulfate without the necessity of doing platelet counts. Daily hypertransfusions of the mice to three to four times normal platelet levels for 4-5 days significantly reduced platelet uptake of radiosulfate to an average of about 40% of the nontransfused controls. Mice rendered thrombocytopenic 48 hr earlier by antiplatelet serum, had 2-day 35S uptakes over 2 1/2 times the controls and 6 times the hypertransfused animals. The administration of a total of 2 ml of serum, given twice daily for 3 days from a thrombocytopenic patient with Hodgkin's disease caused a highly significant 103% rise in radiosulfate incorporation when compared with saline in the hypertransfused mouse. Normal human serum from a healthy donor caused a small and insignificant rise. The serum from a patient with Hodgkin's disease caused a highly significant 63% rise in 35S incorporation when compared to the normal serum.

State of iron(III) in normal human serum: low molecular weight and protein ligands besides transferrin

1970 ◽  
Vol 48 (12) ◽  
pp. 1339-1350 ◽  
Author(s):  
Bibudhendra Sarkar
Keyword(s):  
Molecular Weight ◽  
Human Serum ◽  
Binding Capacity ◽  
Normal Serum ◽  
Normal Human Serum ◽  
Void Volume ◽  
Antibody Concentration ◽  
Low Molecular Weight ◽  
Protein Ligands ◽  
Normal Human

A fraction of Fe(III) in normal human serum is bound to both low molecular weight as well as protein ligands besides transferrin. Citrate was shown to be the major Fe(III)-binding substance in the low molecular weight fraction. Amino acids, sugars, and organic acids, such as ascorbate, pyruvate, and lactate, showed very little or no binding to Fe(III) in normal serum. Iron(III)-binding proteins other than transferrin were shown to be present in normal serum when the native serum with [59Fe(III)] was fractionated by (NH4)2SO4 and Sephadex G-150. The presence of these proteins was observed when trace amounts of Fe(III) were added to the normal serum and when the iron-binding capacity was saturated with Fe(III) to 50% and 100%. These proteins were eluted in the void volume of Sephadex G-150 and none of them corresponded electrophoretically to transferrin. The results of the gel filtration of a mixture of [131I]-transferrin and the proteins eluted in the void volume of Sephadex G-150 were strongly in favor of the Fe(III)-proteins as being neither transferrin aggregates nor transferrin adducts with other proteins. Immunoelectrophoresis of the Sephadex G-150 void volume proteins on agar gel against the antibody to transferrin revealed the absence of transferrin. The presence of at least six proteins in this fraction was shown by immunoelectrophoresis. Positive precipitin reactions were obtained with the antibodies to α2-macroglobulin, γG-globulin, γA-globulin, and γM-globulin. At least two more proteins in this fraction remained unidentified. When the same fraction containing [59Fe(III)] was treated with the whole antisera and the precipitates were counted for radioactivity, a typical antigen-antibody reaction curve was obtained as the antibody concentration was increased. Similar experiments with this fraction and antibodies to α2-macroglobulin, γG-globulin, γA-globulin, and γM-globulin failed to show any significant radioactivity in the precipitate. Since this fraction did not contain any transferrin, it was concluded that there are proteins besides transferrin which can act as ligands for Fe(III) in normal blood plasma.

BIOCOLLOIDS IN NORMAL HUMAN URINE: I. AMOUNT AND ELECTROPHORETIC CHARACTERISTICS

1958 ◽  
Vol 36 (1) ◽  
pp. 1159-1166 ◽  
Author(s):  
T. Webb ◽  
B. Rose ◽  
A. H. Sehon
Keyword(s):  
Human Serum ◽  
Electrophoretic Mobility ◽  
Human Urine ◽  
Filter Paper ◽  
Radioactive Iodine ◽  
Normal Serum ◽  
Γ Globulins ◽  
Normal Human ◽  
Normal Urine ◽  
Serum Components

The biocolloids of normal urine have been isolated and characterized by free electrophoresis and electrophoresis on filter paper. An average of 133 mg of material was recovered from 24-hour aliquots of normal urine. This material was composed of at least seven components as revealed by free electrophoresis at pH 8.6. Five of these components were similar in electrophoretic mobility to the five serum components. A relatively large amount of material was present which behaved like the acid mucoproteins of normal serum. No lipoproteins were detected. Some of the components of the urinary biocolloids were shown to be derived from human serum γ-globulins by labelling the latter with radioactive iodine.

scholarly journals Group IIA-Secreted Phospholipase A2in Human Serum Kills Commensal but Not ClinicalEnterococcus faeciumIsolates

2018 ◽  
Vol 86 (8) ◽  
Author(s):  
Fernanda L. Paganelli ◽  
Helen L. Leavis ◽  
Samantha He ◽  
Nina M. van Sorge ◽  
Christine Payré ◽  
...  
Keyword(s):  
Human Serum ◽  
Normal Serum ◽  
Gram Negative Bacteria ◽  
Opportunistic Pathogens ◽  
Gram Positive ◽  
Content Type ◽  
Gram Positive Bacteria ◽  
Normal Human ◽  
Direct Killing ◽  
The Complement System

ABSTRACTHuman innate immunity employs cellular and humoral mechanisms to facilitate rapid killing of invading bacteria. The direct killing of bacteria by human serum is attributed mainly to the activity of the complement system, which forms pores in Gram-negative bacteria. Although Gram-positive bacteria are considered resistant to killing by serum, we uncover here that normal human serum effectively killsEnterococcus faecium. Comparison of a well-characterized collection of commensal and clinicalE. faeciumisolates revealed that human serum specifically kills commensalE. faeciumstrains isolated from normal gut microbiota but not clinical isolates. Inhibitor studies show that the human group IIA secreted phospholipase A2 (hGIIA), but not complement, is responsible for killing of commensalE. faeciumstrains in human normal serum. This is remarkable since the hGIIA concentration in “noninflamed” serum was considered too low to be bactericidal against Gram-positive bacteria. Mechanistic studies showed that serum hGIIA specifically causes permeabilization of commensalE. faeciummembranes. Altogether, we find that a normal concentration of hGIIA in serum effectively kills commensalE. faeciumand that resistance of clinicalE. faeciumto hGIIA could have contributed to the ability of these strains to become opportunistic pathogens in hospitalized patients.

scholarly journals PRIMARY SERUM TOXICITY AS DEMONSTRATED BY THE CHICKEN EMBRYO

1935 ◽  
Vol 61 (4) ◽  
pp. 489-499 ◽  
Author(s):  
Ernst Witebsky ◽  
Erwin Neter
Keyword(s):  
Guinea Pig ◽  
Human Serum ◽  
Test Object ◽  
Chicken Embryo ◽  
Normal Serum ◽  
Vascular Network ◽  
Inhibitory Influence ◽  
Suitable Test ◽  
Forssman Antigen ◽  
Normal Human

1. The 3 day old chicken embryo removed from its shell is a suitable test object for the demonstration of primary serum toxicity. Addition of normal rabbit type sera as well as Forssman antiserum causes the vascular network to contract and the embryo sinks in the yolk and dies. 2. Only sera of animals of the so called rabbit type produce this phenomenon. Sera of the guinea pig type are ineffective. 3. Heating to 51°C. destroys the complement content of normal human serum as also its effectiveness to produce the vascular phenomenon. 4. Up to the present it has not been possible to reactivate heat-inactivated normal serum by the addition of complement, while inactivated Forssman antiserum can be easily reactivated. 5. The vascular phenomenon of the chicken embryo is produced not only by the addition of a mixture of Forssman antiserum and complement but also by separate addition of both components. 6. Guinea pig type sera, containing dissolved Forssman antigen, are not only ineffective but actually exert an inhibitory influence on effective rabbit type sera as well as on Forssman antiserum.

scholarly journals Group IIA secreted phospholipase A2in human serum kills commensal but not clinicalEnterococcus faeciumisolates

2018 ◽  
Author(s):  
Fernanda L. Paganelli ◽  
Helen L. Leavis ◽  
Samantha He ◽  
Nina M. van Sorge ◽  
Christine Payré ◽  
...  
Keyword(s):  
Human Serum ◽  
Enterococcus Faecium ◽  
Opportunistic Infections ◽  
Normal Serum ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Serum Killing ◽  
Normal Human

AbstractHuman innate immunity employs cellular and humoral mechanisms to facilitate rapid killing of invading bacteria. The direct killing of bacteria by human serum is mainly attributed to the activity of the complement system that forms pores in Gram-negative bacteria. Although Gram-positive bacteria are considered resistant to serum killing, we here uncover that normal human serum effectively killsEnterococcus faecium.Comparison of a well-characterized collection of commensal and clinicalE. faeciumisolates revealed that human serum specifically kills commensalE. faeciumstrains isolated from normal gut microbiota, but not clinical isolates. Inhibitor studies show that the human group IIA secreted phospholipase A2 (hGIIA), but not complement, is responsible for killing of commensalE. faeciumstrains in human normal serum. This is remarkable since hGIIA concentrations in ‘non-inflamed’ serum were considered too low to be bactericidal against Gram-positive bacteria. Mechanistic studies showed that serum hGIIA specifically causes permeabilization of commensalE. faeciummembranes. Altogether, we find that a normal serum concentration of hGIIA effectively kills commensalE. faeciumand that hGIIA resistance of clinicalE. faeciumcould have contributed to the ability of these strains to become opportunistic pathogens in hospitalized patients.ImportanceHuman normal serum contains antimicrobial components that effective kill invading Gram-negative bacteria. Although Gram-positive bacteria are generally considered resistant to serum killing, here we show that normal human effectively kills the Gram-positiveEnterococcus faeciumstrains that live as commensals in the gut of humans. In contrast, clinicalE. faeciumstrains that are responsible for opportunistic infections in debilitated patients are resistant against human serum. The key factor in serum responsible for killing is group IIA secreted phospholipase A2 (hGIIA) that effectively destabilizes commensalE. faeciummembranes. We believe that hGIIA resistance by clinicalE. faeciumcould have contributed to the ability of these strains to cause opportunistic infections in hospitalized patients. Altogether, understanding mechanisms of immune defense and bacterial resistance could aid in further development of novel anti-infective strategies against medically important multidrug resistant Gram-positive pathogens.

scholarly journals Transformation ofLeishmania mexicanametacyclic promastigotes to amastigote-like forms mediated by binding of human C-reactive protein

Parasitology ◽  
2001 ◽  
Vol 122 (5) ◽  
pp. 521-529 ◽  
Author(s):  
A. BEE ◽  
F. J. CULLEY ◽  
I. S. ALKHALIFE ◽  
K. B. BODMAN-SMITH ◽  
J. G. RAYNES ◽  
...  
Keyword(s):  
Human Serum ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Normal Serum ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Metacyclic Promastigotes ◽  
Normal Human ◽  
Kinetics Of

Infective metacyclic promastigote forms ofLeishmania mexicanaare introduced by the bite of sandfly vectors into their human hosts where they transform into the amastigote form. The kinetics of this process was examinedin vitroin response to different combinations of temperature (26 °C or 32 °C), pH (7.2 or 5.5), and exposure to human serum. Little transformation occurred at 26 °C/pH 7.2, intermediate levels at 26 °C/pH 5.5 and 32 °C/ pH 7.2, and the greatest response at 32 °C/pH 5.5. Transformation was stimulated by exposure to normal human serum, but was markedly reduced when serum previously incubated at 56 °C for 1 h was used (complement heat-inactivated). This stimulatory effect was reproduced by exposure to a single purified component of human serum, C-reactive protein (CRP). Binding of CRP to the whole surface ofL. mexicanametacyclic promastigotes, including the flagella, was demonstrated by an indirect fluorescent antibody test. The effect of purified CRP was dose dependent and occurred using normal serum concentrations. The stimulatory effect of whole serum was oblated by CRP depletion and restored by addition of purified CRP. The effects of cAMP analogues indicated that transformation could be mediated via an adenylate cyclase cascade.

scholarly journals AN ANALYSIS OF THE OPSONIC AND TROPIC ACTION OF NORMAL AND IMMUNE SERA BASED ON EXPERIMENTS WITH THE PNEUMOCOCCUS

1933 ◽  
Vol 57 (4) ◽  
pp. 527-547 ◽  
Author(s):  
Hugh K. Ward ◽  
John F. Enders
Keyword(s):  
Human Serum ◽  
Immune Serum ◽  
Normal Serum ◽  
Heat Stable ◽  
Prolonged Contact ◽  
Current Usage ◽  
Single Well ◽  
Predominant Factor ◽  
Normal Human ◽  
Experimental Difficulty

1. In normal unheated human serum, virulent pneumococci may be prepared for phagocytosis by two separate antibodies, acting in conjunction with complement. One of these is the type-specific anticarbohydrate antibody reacting with the carbohydrate fraction of the pneumococcus. The other is probably also a type-specific antibody, but quite distinct from the former, and therefore must react with a different antigenic constituent of the bacterium. 2. In the normal human serum heated to 56°C., these two antibodies may, after prolonged contact with the organism, promote phagocytosis of the pneumococcus without the adjuvant action of complement. 3. Although these two antibodies are equally effective in the phagocytosis of 24 hour culture organisms by normal blood, the anticarbohydrate antibody tends to become the predominant factor as the pneumococci approach the state in which they exist in the animal body. 4. In so far as we have been able to show, the anticarbohydrate antibody is the only antibody in immune serum which can induce phagocytosis. This substance by itself is active in a phagocytic system, but just as in the normal serum, complement enhances its effect. The failure to demonstrate the presence in the immune serum of an antibody, distinct from the anticarbohydrate antibody, analogous to that found in the normal serum, may be due to the experimental difficulty of removing all the anticarbohydrate antibody from a concentrated immune serum. 5. Thus it is seen that a single well defined antibody (the anticarbohydrate antibody) may be responsible for the phagocytic action of normal unheated serum, normal heated serum, inactivated immune serum, and immune serum activated by complement. These facts appear to us to invalidate Neufeld's division of the phagocytic antibodies into (a) bacteriotropins (antibodies, the phagocytic titre of which is not raised by the addition of complement); (b) opsonic antibodies (antibodies, comparable to the lysins, which are only active in the presence of complement). 6. Complement alone is incapable of inducing phagocytosis of the pneumococcus. In the phagocytic process, it appears simply to increase the speed at which the reaction takes place. Its role may be compared to that of a catalyst in a chemical reaction. 7. On the basis of these findings, it is proposed that the term "tropin" be discarded as misleading and unnecessary, and that the term "opsonin" be retained to denote any heat-stable antibody which prepares bacteria for phagocytosis. Contrary to current usage, it would not suggest a combination of antibody with complement.

scholarly journals A Study of Non-specific Complement-fixation with particular reference to the Interaction of Normal Serum and certain Non-antigenic substances

1928 ◽  
Vol 28 (2) ◽  
pp. 172-197 ◽  
Author(s):  
T. J. Mackie ◽  
M. H. Finkelstein
Keyword(s):  
Human Serum ◽  
Complement Fixation Test ◽  
Complement Fixation ◽  
Normal Serum ◽  
Fixation Test ◽  
Normal Human Serum ◽  
Rabbit Serum ◽  
Considerable Proportion ◽  
Normal Human ◽  
Fixation Reaction

1. When a solution of commercial peptone is substituted for antigen in a complement-fixation test with the unheated normal serum of certain species (man, ox, sheep, horse, rabbit, white rat), a definite fixation reaction occurs both at 37° C. and at 0° C. In the ox, sheep, horse and rabbit this property of serum is partially stable at 55° C., but normal human serum and the serum of the white rat are inactive after heating at this temperature. The property is resident mainly in the carbonic-acid-insoluble globulins of the serum.2. The same results are obtained when ethyl alcohol diluted with several volumes of normal saline solution is substituted for antigen in a complement-fixation test with normal serum.3. Analysis of these reactions shows a close correspondence with complement-fixation by the sera of normal animals plus the Wassermann “antigen”—the Wassermann reaction of normal animals.4. Marked complement-fixation effects are also obtained with heated normal serum of the rabbit, ox, sheep, horse plus cholesterol suspension, and particularly cholesterolised-peptone, these effects occurring in parallel with those produced by serum plus alcohol-saline, peptone solutions and the Wassermann “antigen.” The heated normal serum of the pig, white rat and guinea-pig do not exhibit these reactions, and the same applies to heated normal human serum. Unheated pig serum fails to react. Such results also elicit a close relationship between these non-specific reactions and the Wassermann reactions of normal animals.5. The reacting property is absent from the serum (heated and unheated) of young rabbits during the first 2 to 3 weeks of life, but appears soon after this (e.g. by the 37th day) and is progressive in development. Its development in early life runs parallel to that of the natural haemolytic property of the serum for sheep's blood (due to a natural antibody-like substance). The two properties are, however, independent as illustrated by absorption tests.6. Besides the agents referred to above as capable of fixing complement along with normal sera, other substances possess a similar property, e.g. certain alcohols, sodium oleate, tissue proteins, certain amino-acids and sodium nucleate. Commercial peptone purified by precipitation with alcohol is equally active with the original material. Cholesterolisation of these agents may yield a product whose activity is greater than that due to summation of effects.7. Wassermann-positive and -negative human sera have been tested in the complement-fixation reaction with certain of these “pseudo-antigens,” viz. alcohol-saline, peptone, cholesterol, and cholesterolised-peptone, but a uniform parallelism has not been demonstrated between the reactions with these agents and the Wassermann effect. Some Wassermann-positive sera react also with alcohol-saline, peptone, cholesterol and cholesterolised-peptone, while sera from selected normal persons are quite inactive. A considerable proportion of Wassermann-positive sera yields definite complement-fixation with cholesterol and cholesterolised-peptone; a small proportion of Wassermann-negative sera reacts with these agents.8. The thermolability of the serum principles acting with various “pseudoantigens” has been studied by testing unheated serum and serum heated at temperatures ranging from 46° to 60° C. Two types of thermolability curve have been demonstrated with different specimens of rabbit serum: (1) a more or less progressive weakening of the various reactions with inactivation at 60° C.; (2) inactivation of the effects with Wassermann “antigen,” alcoholsaline and cholesterol at 50–52° C., activation of the effects with the Wassermann “antigen” and cholesterol at 54–56°C. and inactivation again above 60° C.; in this case the curves for peptone and cholesterolised-peptone do not show such double inactivation. Unheated normal human serum yields reactions with the various agents (including the Wassermann “antigen”) but inactivation occurs at 50° to 54° C. whereas certain syphilitic sera yield thermolability curves somewhat similar to type (1) of rabbit serum, with inactivation at 60° C. or over.

scholarly journals DANGEROUS UNIVERSAL DONORS

Blood ◽  
1950 ◽  
Vol 5 (6) ◽  
pp. 553-567 ◽  
Author(s):  
DONALD M. ERVIN ◽  
RICHARD M. CHRISTIAN ◽  
LAWRENCE E. YOUNG
Keyword(s):  
Human Serum ◽  
Whole Blood ◽  
Normal Human Serum ◽  
Random Group ◽  
A Cells ◽  
Group A ◽  
Normal Human ◽  
Screening Procedures

Abstract 1. Severe hemolytic reactions were observed in 3 group A (subgroup A1) recipients transfused with group O whole blood or plasma. In one case, 10 ml. of a commerical preparation of soluble A and B factors had been added to 500 ml. of whole blood prior to the transfusion and it is believed that the reaction might have been even more serious had not this material been added. 2. The anti-A antibodies in the serum of the dangerous universal donors causing the hemolytic reactions fixed complement, acted as hemolysins, were difficult to neutralize with soluble A and B factors, were capable of giving positive Coombs tests and their ability to agglutinate A cells was enhanced by the presence of normal human serum. These characteristics were similar to those observed in serum from donors known to be actively immunized against the A factor, but the stimulus for development of "immune" anti-A antibodies in the dangerous group O donors was not apparent. 3. Small amounts of immune A antibody were consistently demonstrated in 12 of 100 random group O sera which, after neutralization, produced indirect Coombs tests with A1 cells and agglutinated A1 cells suspended in compatible normal human serum. 4. Screening procedures for elimination of dangerous group O donors are discussed.

scholarly journals The inhibitory action upon subsequent Phagocytosis, exerted on active normal serum by inactive normal serum through which Bacilli have been passed

1907 ◽  
Vol 79 (534) ◽  
pp. 482-484 ◽  
Keyword(s):  
Human Serum ◽  
Salt Solution ◽  
Inhibitory Action ◽  
Normal Serum ◽  
Normal Human Serum ◽  
The Other ◽  
Supernatant Fluid ◽  
Other Hand ◽  
Difference Of Opinion ◽  
Normal Human

Considerable difference of opinion still prevails regarding the nature of the opsonic substances present in normal serum. Wright, Bulloch and Atkin, etc., uphold the view that the opsonin of normal serum is a simple thermolabile body. Muir, on the other hand, regards the opsonin as a body which behaves like complement, while Dean holds that it is essentially thermostable and in all probability co-operates in its action with a thermolabile complement. The demonstration of anti-bodies by complement-deviation experiments (Bordet, Gengou, Pfeiffer and Friedberger, etc.) has recently proved fruitful in connection with the bacteriolysins, hæmolysins, precipitins of immune sera, and the following experiments were designed to test whether, by a similar method applied to phagocytosis, the presence in normal serum of opsonic amboceptors could be demonstrated :─ Experiment I. Normal human serum was heated for 30 mins. at 60° C. (denoted “ A ”). A very thick emulsion of tubercle bacilli in 1 : 1000 salt solution was added in equal volumes to “ A ” and kept in contact therewith for 1 hr. 30 mins. at 37° C. The mixture was then centrifugalised (7000 revolutions per minute) for 1 hr., and the supernatant fluid pipetted off (denoted “ B ”).

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