scholarly journals FURTHER STUDY ON THE CULTURAL CONDITIONS OF LEPTOSPIRA (SPIROCHÆTA) ICTEROHÆMORRHAGIÆ

1918 ◽  
Vol 27 (5) ◽  
pp. 593-608 ◽  
Author(s):  
Hideyo Noguchi
Keyword(s):  
Guinea Pig ◽  
Human Serum ◽  
Salt Solution ◽  
Cultural Value ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Ringer’S Solution ◽  
Moderate Growth ◽  
Nutrient Value ◽  
Isotonic Salt Solution

1. The presence of suitable animal or human serum is essential for the cultivation of Leptospira icterohæmorrhagiæ. 2. The nutrient value of serum is considerably reduced by heating to 60°C. for 30 minutes and is destroyed by boiling (100°C). Filtration through a Berkefeld filter does not diminish the nutrient value of the serum. 3. The cultural value of different animal sera varies considerably. It is entirely absent from the sera of the rat and the pig. The sera of the rabbit, horse, and goat are better suited for the growth of the organism than those of the guinea pig, sheep, donkey, or calf. Human serum is suitable, but not ascitic fluid. 4. Fresh or heated emulsions of the liver, kidney, heart muscle, or testicle of the normal guinea pig or rabbit have no cultural value for the organism. The same may be said of both the white and yolk of the hen's egg. 5. A luxuriant growth takes place in a medium of Ringer's solution to which more than 10 per cent of normal rabbit serum is added. There is only moderate growth with 5 per cent of serum, and none when less than 2 per cent is present. The use of an undiluted serum offers no advantage over a diluted one, provided the latter contains at least 10 per cent of serum. In the case of certain animal sera dilution seems to make them more suitable for cultivation purposes, owing perhaps to its reduction of their inherent alkalinity. 6. The tonicity of the culture medium has but little influence upon the growth and morphology of the organism. A medium containing distilled water as diluent or one containing 8 per cent sodium chloride seems to give identical results. The viability of the organism was greatest in a medium in which Ringer's solution or isotonic salt solution was used as diluent. 7. The reaction of the medium is an important factor in the cultivation of the organism, which thrives most vigorously in a medium of which the reaction is slightly alkaline, not exceeding that of the serum. If the reaction is neutral, the growth is meager, and the culture is short lived. When the reaction of a medium becomes alkaline by the addition of a small amount of sodium hydroxide, or faintly acid by the addition of a little hydrochloric acid, no growth can take place. 8. Leptospira icterohæmorrhagiæ is an obligatory aerobe. Any hindrance to the access of oxygen constitutes an unfavorable factor in obtaining a culture. 9. The addition of carbohydrates to media has no perceptible effect upon the growth or morphology of the organism. The reaction of the media is not modified by their presence. 10. Leptospira icterohæmorrhagiæ grows at any temperature between 37° and 10°C., the optimum zone being 30–37°C. Growth proceeds more rapidly at 37°C. than at 30° or at 25°, but the cultures remain viable much longer at the latter temperatures. No growth takes place at 42°C. 11. Three different media are described for the cultivation of freshly isolated strains. After prolonged cultivation on these media a strain may be readily cultivated in a serum diluted with Ringer's or isotonic salt solution.

scholarly journals STUDIES ON THE MECHANISM OF THE FORMATION OF THE PENICILLIN ANTIGEN

1961 ◽  
Vol 114 (6) ◽  
pp. 875-940 ◽  
Author(s):  
Bernard B. Levine ◽  
Zoltan Ovary
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Penicillin G ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Beings ◽  
Mixed Disulfide ◽  
Diastereomeric Mixture ◽  
Potassium Penicillin

An excess of D-benzylpenicillenic acid (BPE) was reacted with human γ-globulin, human serum albumin, gelatin, and poly-L-lysine in aqueous solution buffered at pH 7.5–8.0. Under these conditions, BPE reacted predominantly with lysine ϵ-amino groups of the proteins to form the mixture of diastereomers of ϵ-N-(D-α-benzylpenicilloyl)-lysine groups (Di-BPO-Lys). BPE reacted also, but to a considerably smaller extent, with cystine disulfide linkages of human γ-globulin and human serum albumin to form D-benzylpenicillenic acid-cysteine mixed disulfide groups (BPE-SS-Cys). Conjugates containing large numbers of BPE or D-penicillamine mixed disulfide groups were prepared by reaction of BPE or D-penicillamine with thiolated human γ-globulin under mild oxidizing conditions. Anti-penicillin antibodies were produced in rabbits by immunization with either potassium penicillin G (PG) or a preincubated mixture of PG with normal rabbit serum (PG-NRS) in complete Freund's adjuvant. Specific precipitation analyses in aqueous and gel media (Ouchterlony), PCA analyses, and specific inhibition of these reactions with haptens were carried out on the rabbit anti-PG and anti-(PG-NRS) sera, using the above conjugates as antigens. The anti-penicillin antibodies were found to be directed against the diastereomeric mixture of N-(D-α-benzylpenicilloyl) groups, predominantly the Di-BPO-Lys groups. By these techniques, no antibodies directed against the BPE-mixed disulfide or the D-penicillamine mixed disulfide groups were detected. Three out of six patients with histories of allergic reactions to PG responded with wheal-and-erythema reactions to the N-(D-α-benzylpenicilloyl) (BPO) groups contained in BPE-human gamma globulin conjugate. Another such patient exhibited serum antibodies specific for the BPO group. One patient being treated with 25 gm per day of PG showed the presence of non-dialyzable antigenic BPO-conjugates in his serum. These results demonstrate that the diastereomeric BPO groups (predominantly Di-BPO-Lys groups) are major antigenic determinant groups responsible for PG hypersensitivity in rabbits and human beings. The possible clinical usefulness of multivalent Di-BPO conjugates and univalent Di-BPO haptens is discussed.

BIOTRANSFORMATION OF PREGNENOLONE AND PROGESTERONE BY RABBIT OVARIAN FOLLICLES AND CORPORA LUTEA

1973 ◽  
Vol 74 (4) ◽  
pp. 775-782 ◽  
Author(s):  
Edward V. YoungLai
Keyword(s):  
Granulosa Cells ◽  
Salt Solution ◽  
Hydroxysteroid Dehydrogenase ◽  
Ovarian Follicles ◽  
Rabbit Serum ◽  
Corpora Lutea ◽  
Normal Rabbit Serum ◽  
Theca Cells ◽  
Theca Interna

ABSTRACT Rabbit ovarian follicles obtained prior to and after mating and corpora lutea (24 h and 48 h post-coitus) were incubated with radioactive pregnenolone and progesterone to determine whether these substrates could serve as precursors of androgens and oestrogens. Incubations were carried out for 3 h in Hanks balanced salt solution: medium 199: normal rabbit serum: 55:30:15. Granulosa cells and corpora lutea converted pregnenolone to progesterone but no labelled oestrogens could be detected. Trace amounts of androgens were synthesized by the granulosa cells. Whole sliced follicles and theca formed progesterone from pregnenolone and androgens from both substrates. Oestradiol-17β was only synthesized by the whole sliced follicles and in one experiment by theca cells. Mating caused an increase in the 3β-hydroxysteroid dehydrogenase for pregnenolone and formation of testosterone but no oestrogens by the theca cells. These results indicate that both theca interna and granulosa cells are needed for oestrogen biosynthesis by rabbit follicles.

scholarly journals Humoral immunostimulation. IV. Role of complement.

1975 ◽  
Vol 141 (4) ◽  
pp. 736-752 ◽  
Author(s):  
W T Shearer ◽  
J P Atkinson ◽  
M M Frank ◽  
C W Parker
Keyword(s):  
Cell Growth ◽  
Guinea Pig ◽  
Human Serum ◽  
Calf Serum ◽  
Rabbit Serum ◽  
Mouse Serum ◽  
Low Concentrations ◽  
High Concentrations ◽  
Nucleoside Uptake ◽  
Pig Serum

When L cells were treated with anti-L-cell antibody in medium containing heat-inactivated fetal calf serum, nucleoside uptake and cell growth were stimulated. The response was markedly increased when fresh, unheated sera from calves, guinea pigs, humans, mice, or rabbits were also present. The factors in unheated serum responsible for the enhancement of immunostimulation were studied. Using low concentrations of sera deficient in various complement (C) components and low concentrations of antibody no augmentation of immunostimulation was seen with Clr-deficient human serum, C2-deficient human serum, C2,4-deficient human serum, C4-deficient guinea pig serum, C3-C9-depleted guinea pig serum (by administration of cobra venom factor to animals), but stimulation was observed with C5-deficient human serum, C5-deficient mouse serum, and C6-deficient rabbit serum. When the concentration of anti-serum was raised, however, augmentation was observed with C4-deficient guinea pig serum. Thus, at low concentrations of antiserum enhancement appeared to occur through the classical C pathway, whereas at high concentrations of antibody either the classical or alternate C pathways appeared to be involved. Stimulation was specifically restored by purified C2 in C2-deficient serum and by C3 in C3-C9-deficient serum. Under the usual reaction conditions consumption of guinea pig C component C4 could be demonstrated which provided direct evidence for activation of the classical C pathway under conditions leading to immunostimulation. By immunofluorescence, cells treated with antibody and normal human serum had human C3 deposited at the cell surface. Taken together these observations suggest that C activated through C3 by either the classical or alternate pathways has the potential to enhance nucleoside incorporation into DNA and cell growth of cells exposed to limiting amounts of antibody. Although the mechanism of stimulation is unknown, it is likely to involve a direct effect of C3 at the level of the cell membrane.

scholarly journals THE TOXICITY OF HUMAN SERUM FOR THE GUINEA PIG

1929 ◽  
Vol 49 (3) ◽  
pp. 497-506 ◽  
Author(s):  
Susan Griffith Ramsdell ◽  
I. Davidsohn
Keyword(s):  
Guinea Pig ◽  
Human Serum ◽  
Rabbit Serum ◽  
Differential Absorption ◽  
Right Heart ◽  
Circulatory Effects ◽  
The Right ◽  
Heterophilic Antibody ◽  
Immune Rabbit

From the foregoing study there are indications: 1. Of toxicity in all fresh human serum for the guinea pig; this toxicity tends to disappear after 48 hours after bleeding, and its manifestations are strikingly similar to those of the heterophilic antibody in immune rabbit serum. 2. Of an increased toxicity in the serum of antisera-treated human cases; this is usually coexistant with the production of other antibodies; it tends likewise to disappear in time after treatment; differential absorption experiments indicate that its character is heterophilic, and its manifestations differ from those of anaphylaxis in that certain circulatory effects—hemorrhage and increased edema in the lungs and distension of the right heart—are added to the usual findings in true anaphylactic deaths.

scholarly journals A STUDY OF THE EFFECT OF SERUM ON THE IMMUNOLOGICAL REACTION OF A BACTERIAL ENDOTOXIN

1956 ◽  
Vol 103 (4) ◽  
pp. 439-452 ◽  
Author(s):  
Leighton E. Cluff
Keyword(s):  
Human Serum ◽  
Serratia Marcescens ◽  
Normal Serum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Immunological Reaction ◽  
Beta Globulin ◽  
Zone Electrophoresis ◽  
Polysaccharide Antigens ◽  
Relationship Of

Contact of a purified endotoxin from Shigella fiexneri Type Z with normal rabbit or human serum results in an alteration of its immunological reaction with antiserum, as determined by precipitation in gel. Analysis of fractions of normal serum obtained by zone electrophoresis in starch indicates that the component responsible for altering the immunological reaction of endotoxin is associated with beta globulin. Normal serum has no similar effect on the immunological reaction of a variety of other protein and polysaccharide antigens. Serum from rabbits made tolerant to the pyrogenic action of an endotoxin from Serratia marcescens (P-35) possesses the ability to alter the reaction of Shigella endotoxin with its specific antiserum, although the serum from tolerant rabbits does not significantly enhance the pyrogenicity of Shigella toxm. The component of normal rabbit serum responsible for the effect on the immunological reaction of Shigella endotoxin is not destroyed by heating at 56°C. The possible relationship of the effect of normal serum on the immunological reaction of endotoxin and the augmentation of fever induced by endotoxins is discussed.

Identification of Maize Rayado Fino Virus by Immunoelectron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 636-637
Author(s):  
O. E. Bradfute
Keyword(s):  
Electron Microscopy ◽  
Zea Mays ◽  
Costa Rica ◽  
Severe Disease ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Virus Particles ◽  
Far North ◽  
Maize Rayado Fino Virus ◽  
Antibody Coating

Maize rayado fino virus (MRFV) causes a severe disease of corn (Zea mays) in many locations throughout the neotropics and as far north as southern U.S. MRFV particles detected by direct electron microscopy of negatively stained sap from infected leaves are not necessarily distinguishable from many other small isometric viruses infecting plants (Fig. 1).Immunosorbent trapping of virus particles on antibody-coated grids and the antibody coating or decoration of trapped virus particles, was used to confirm the identification of MRFV. Antiserum to MRFV was supplied by R. Gamez (Centro de Investigacion en Biologia Celular y Molecular, Universidad de Costa Rica, Ciudad Universitaria, Costa Rica).Virus particles, appearing as a continuous lawn, were trapped on grids coated with MRFV antiserum (Fig. 2-4). In contrast, virus particles were infrequently found on grids not exposed to antiserum or grids coated with normal rabbit serum (similar to Fig. 1). In Fig. 3, the appearance of the virus particles (isometric morphology, 30 nm diameter, stain penetration of some particles, and morphological subunits in other particles) is characteristic of negatively stained MRFV particles. Decoration or coating of these particles with MRFV antiserum confirms their identification as MRFV (Fig. 4).

Localization of oxytocin and neurophysin in baboon (Papio anubis) corpus luteum by immunocytochemistry

1986 ◽  
Vol 113 (4) ◽  
pp. 570-575 ◽  
Author(s):  
Firyal S. Khan-Dawood
Keyword(s):  
Corpus Luteum ◽  
Rabbit Serum ◽  
Corpora Lutea ◽  
Positive Staining ◽  
Normal Rabbit Serum ◽  
Fallopian Tubes ◽  
Papio Anubis ◽  
Luteal Cells ◽  
Luteal Tissue ◽  
Immunocytochemical Reaction

Abstract. Immunoreactive oxytocin is detectable in the corpora lutea of women and cynomolgus monkeys by radioimmunoassay. To localize the presence of oxytocin and neurophysin I in ovarian tissues of subhuman primates, three corpora lutea and ovarian stromal tissues and two Fallopian tubes obtained during the menstrual cycle of the baboon and decidua from two pregnant baboons were examined using highly specific antisera against either oxytocin or neurophysin I and preoxidase-antiperoxidase light microscopy immunohistochemistry. Oxytocin-like as well as neurophysin I-like immunoreactivities were found in some cells of all the corpora lutea only, but could not be demonstrated in ovarian stromal tissues, Fallopian tubes and decidua. Specificity of the immunocytochemical reaction was further confirmed by immunoabsorption of the antiserum with excess oxytocin or neurophysin, after which the immunoreactivities for both oxytocin and neurophysin in the luteal tissue were negative. Similar controls using normal rabbit serum gave no positive staining for either oxytocin or neurophysin. Counterstaining of the positive immunoreactivities for oxytocin and neurophysin I with Mayer's haematoxylin and eosin demonstrated clearly that the oxytocin and neurophysin I appeared as granular material mainly within the cytoplasm of the luteal cells. The localization of immunoreactive oxytocin and neurophysin I in the corpus luteum of the baboon demonstrates directly the presence of these two neurohypophysial peptides within primate luteal cells and suggests their local production.

scholarly journals Interactions of C-reactive protein with the complement system. III. Complement-dependent passive hemolysis initiated by CRP.

1975 ◽  
Vol 142 (5) ◽  
pp. 1065-1077 ◽  
Author(s):  
A.P. Osmand ◽  
R.F. Mortensen ◽  
Joan Siegel ◽  
H. Gewurz
Keyword(s):  
Guinea Pig ◽  
Human Serum ◽  
Complete System ◽  
Gel Filtration ◽  
C Reactive Protein ◽  
Inflammatory Reactions ◽  
Reactive Protein ◽  
Rabbit Igg ◽  
The Complement System ◽  
Pig Serum

Interactions of CRP with various substrates in the presence of human serum have been shown to result in efficient activation of C components C1-C5. We now report the ability of CRP to initiate C-dependent hemolysis. For this purpose CRP was isolated by affinity chromatography using pneumococcal CPS and gel filtration; its purity was established by several criteria. Erythrocytes were coated with CPS (E-CPS) and passively sensitized with CRP. C-dependent lysis of these cells was observed upon the addition of suitably absorbed human serum, and the efficiency of hemolysis compared favorably with that initiated by rabbit IgG anti-CPS antibody. CRP also sensitized E-CPS for lysis by guinea pig C; partial lysis was seen when C4-deficient guinea pig serum was used, suggesting that CRP also shares with antibody the ability of CRP to fully activate the C system and provide further evidence for a role for CRP similar to that of antibody in the initiation and modulation of inflammatory reactions via the complete system.

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