bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation and Characterization of a Protective Antigen-Containing Particle from Culture Supernatant Fluids of
Erysipelothrix rhusiopathiae
Authors: White R. R. , Verwey W. F.
Published: January 2020

The mouse-protective activity of
Erysipelothrix rhusiopathiae
culture supernatant fluids exists in a polydisperse form, ranging in density from aggregates which sediment at 10,000 ×
g
for 3 hr to soluble units which will not sediment at 198,000 ×
g
for 12 hr. A partially purified protective antigen has been isolated from the aggregates sedimented from a concentrate of the culture supernatant fluid at 20,000 ×
g
for 3 hr. These aggregates contained the major protective antigen or antigens of
E. rhusiopathiae
, since, in addition to inducing active immunity, they adsorbed essentially all of the passively protecting antibody from rabbit antiserum produced by immunization with whole culture. The protective activity in these aggregates was destroyed by trypsin and greatly diminished by muramidase and heating at 64 C, but was not affected by lipase or ribonuclease.
Dependence of Time to Death on Molecular Size of Botulinum Toxin
Authors: Lamanna Carl , Spero Leonard , Schantz Edward J.
Published: January 2020

The molecular size of type A botulinum toxin affects the response in a time-to-death assay. Definitive statements of specific activity should be based on quantal assay.
Subcellular Localization of
Salmonella enteritidis
Endotoxin in Liver and Spleen of Mice and Rats
Authors: Willerson James T. , Trelstad Robert L. , Pincus Theodore , Levy Stuart B. , Wolff Sheldon M.
Published: January 2020

Salmonella enteritidis
14
C-endotoxin was recovered predominantly from the nuclear and mitochondrial subcellular fractions of livers and spleens of mice and rats, 3.5 hr and 3 days after intravenous administration. Of the recovered radioactivity, 10 to 20% was present in the liver mitochondrial fraction as high-molecular-weight, biologically active material, suggesting the presence of intact endotoxin. Autoradiographic studies demonstrated nuclear and cytoplasmic labeling in the liver and at least nuclear label in spleen cells. The resistance of rats, as compared to mice, to the induction of amyloidosis does not appear to be based on a difference in subcellular localization of endotoxin within the reticuloendothelial system.
Interconversion of Purine Mononucleotides in
Pasteurella pestis
Authors: Brubaker R. R.
Published: January 2020

Included among the five established determinants of virulence in
Pasteurella pestis
are the abilities of cells to accomplish the de novo biosynthesis of purines and to grow as dark pigmented (P
+
) colonies on a solid synthetic medium containing hemin. P
+
isolates of
P. pestis
strain KIM-10 (mouse intraperitoneal
ld
50
< 10 cells) failed to convert exogenous guanine-
8
-
14
C
to adenine residues of ribonucleic acid (RNA) when cultivated in a minimal medium which favored the pigmentation reaction. This conversion occurred in P
+
cells grown in an enriched medium which did not support the pigmentation reaction and was observed in P
−
mutants cultivated in both types of media. Both P
+
and P
−
isolates converted exogenous adenine-
8
-
14
C
but not adenine-
2
-
14
C
at a significant rate to guanosine residues of RNA when grown under a variety of conditions. This difference appeared to reflect a deficiency of adenine deaminase. The mouse intraperitoneal
ld
50
of purine-auxotrophs was about 10
2
cells when the metabolic block occurred prior to the de novo formation of inosine monophosphate (IMP). In contrast, the corresponding value for a mutant blocked between IMP and guanine monophosphate was > 10
7
cells in mice and > 10
8
cells in guinea pigs.
Streptolysin O: Activation by Thiols
Authors: Bernheimer Alan W. , Avigad Lois S.
Published: January 2020

Various straight-chain thiols produce approximately the same maximal activation of streptolysin O.
Immunoglobulin Class of Natural Human Antibodies Reactive with
Treponema pallidum
Authors: Julian A. J. , Logan L. C. , Norins L. C. , Cohen I. R.
Published: January 2020

To aid the development of serological tests for incubating syphilis, a characterization was made of the background of natural anti-
Treponema pallidum
antibodies against which the initial immune response to syphilis takes place. Sera from presumed normal persons were studied with monospecific antisera in an indirect fluorescent-antibody procedure. Of 36 sera tested at a 1:5 dilution, all showed IgG reactivity with
T. pallidum
(Nichols strain), 58% showed IgM reactivity, and 20% showed IgA reactivity. The titer of IgG reactivity was considerably higher than that of the other two immunoglobulins. Heating the sera for 1 hr at 65 C abolished IgA and IgM anti-
T. pallidum
reactivity, but one-third of the sera retained IgG reactivity. Human Cohn fractions II and III
1
from three commercial sources contained mostly IgG antibodies reactive with
T. pallidum
, but IgM and IgA antibodies were also present. IgG reactivity was found in a pool of presumably normal maternal sera from 20 mothers and in a corresponding pool of umbilical cord serum from their infants. IgM and IgA reactivities were present only in the maternal serum pool.
Response of sublethally irradiated monkeys to a replicating viral antigen
Authors: Hilmas D E , Spertzel R O
Published: January 2020

Temporal effects of exposure to sublethal, total-body X radiation (400 R) on responses to vaccination with attenuated Venezuelan equine encephalomyelitis vaccine virus. TC-83, were examined in rhesus monkeys. Viremia, often with delayed onset, was prolonged even when irradiation preceded vaccination by 28 days. Virus titers were increased, articularly in groups irradiated 4 or 7 days before vaccination. Delay in appearance of hemagglutination-inhibition and serum-neutralizing antibody correlated closely with persistence of viremia in irradiated-vaccinated monkeys. The temporal course of antibody response was markedly affected by the intervals between irradiation and injection of this replicating antigen. With longer intervals between irradiation and vaccination, the somewhat depressed antibody responses approached normal or surpassed those of nonirradiated monkeys. Vaccination 14 days after radiation exposure resulted in lethality to 8 of 12 monkeys, apparently as a result of secondary infection. The additional lymphopenic stress due to the effect of TC-83, superimposed on the severly depressed hematopoietic competence at 14 days, undoubtedly contributed to this increased susceptibility to latent infection.
Ultrastructural cytochemical evidence for the activation of lysosomes in the cytocidal effect of Chlamydia psittaci
Authors: Todd W J , Storz J
Published: January 2020

The cytopathic effect of the polyarthritis strain of Chlamydia psittaci was studied in cultured bovine fetal spleen cells and found to be mediated by the release of lysosomal enzymes into the host cytoplasm during the late stages of chlamydial development. Ultrastructural cytochemical analysis and cell fractionation studies of infected cells revealed a close relationship between the stage of chlamydial development, fine structural features of the host, and localization of lysosomal enzyme activities. After adsorption, chlamydiae entered the host cells by endocytosis. The endocytic vacuoles containing individual chlamydiae and later the inclusion vacuoles containing the different chlamydial developmental forms were always free from lysosomal enzyme activity. Even after extensive multiplication of chlamydiae, lysosomal enzymes remained localized within lysosomes or their precursors in the host cell. Coincident with the process of chlamydial maturation, lysosomal enzymes were released into the host cytoplasm and were always associated with disintegration of host cell constituents and lysis. The chlamydiae appeared to be protected from this lysosomal enzyme activity by the inclusion membrane. After release from the inclusion, elementary bodies maintained their fine structural features, whereas all other chlamydial developmental forms lost their ultrasturctural integrity.
Guanidine extraction of streptococcal M protein
Authors: Russell H , Facklam R R
Published: January 2020

A new method of extracting M protein from streptococcal cell walls has been presented. The extracting agent was guanidine-hydrochloride, a protein denaturant. The crude guanidine extract was further purified by ammonium sulfate and pH 5 fractionation and by hydroxyapatite column chromatography. Three major protein peaks were eluted from the hydroxyapatite column with 0.01, 0.1 and 0.3 M phosphate buffer, respectively. Protein fractions eluted at 0.1 and 0.3 M phosphate concentractions contained antigens that precipitated with homologous M-protein specific antisera, whereas the 0.01 M phosphate fraction had no immunological activity. The fraction eluted with 0.3 M phosphate was electrophoretically homogeneous in sodium dodecyl sulfate-acrylamide gels and elicited the production of bactericidal antibodies in rabbits. The 0.1 M phosphate buffer eluant was electrophoretically heterogeneous and did not elicit the production of bactericidal antibodies in rabbits.
Biological activities of endotoxins detoxified by alkylation
Authors: Chedid L , Audibert F , Bona C , Damais C , Parant F , Parant M
Published: January 2020

It has been previously observed that lipopolysaccharides can be detoxified by alkylation and yet retain their adjuvant activity. Our present findings confirm these results and show, moreover, that these derivatives did not lose their capacity to protect mice against lethal irradiation and lost only partially their ability to interrupt pregnancy or to induce blast transformation of murine B-lymphocytes. However, in contrast with lipopolysaccharides, these alkylated preparations did not enhance the nonspecific resistance of mice to a bacterial infection. The various bilogical functions of endotoxins can therefore be separated and are not uniformly related to their toxicity.
Nature of "memory" in T-cell mediated antibacterial immunity: cellular parameters that distinguish between the active immune response and a state of "memory"
Authors: North R J , Deissler J F
Published: January 2020

Immunizing infection in mice with Listeria monocytogenes resulted in the generation of two distinct states of immunological reactivity. There was generated (i) a short-lived state of active immunity that functioned to urgently eliminate the infection organism from the tissues and (ii) a long-lives state of increased immunological potential that enabled the host to respond to seconday infection in an accelerated manner. Short-lived active immunity was mediated by replicating T cells and expressed by activated macrophages, and it ended when these cell types disappeared from the tissue soon after complete elimination of the parasite. Long-lived immunological protential was associated with a persistent level of delayed sensitivity and with the presence of a small number of nonreplicating protective T cells. It is suggested that the state of delayed sensitivity represents a state of immunological T-cell memory of the cell-mediated type.
Effect of the quality of the lipopolysaccharide on mouse virulence of Salmonella enteritidis
Authors: Valtonen M V , Plosila M , Valtonen V V , Mäkelä P H
Published: January 2020

The cell wall O antigen is known to affect mouse virulence of Salmonella. We have shown earlier that S. typhimurium expressing somatic antigen 9,12 is less virulent than its 4,12 sister transductants in mice after intraperitoneal inoculation, suggesting that the 4,12-type O antigen is connected with high virulence in mice. In this report we show, in accord with this suggestion, that when the naturally occurring O-9,12 S. enteritidis is made O-4,12 by transduction, its virulence is increased. The difference between O-9,12 and O-4,12 sister transductants is highly significant, with P less than 0.001.
Some properties of a D-alanine carboxypeptidase in envelope fractions of Neisseria gonorrhoeae
Authors: Davis R H , Salton M R
Published: January 2020

Envelope preparations of Neisseria gonorrhoeae strain GC1 (a stable, piliated strain of intermediate colony morphology) and type T1 possess a D-alanine carboxypeptidase which releases the terminal alanine residue from the uridine 5'-diphosphate-N-acetyl muramylpentapeptide substrate (isolated from Bacillus cereus T). The D-alanine carboxypeptidase of the GC1 envelopes has a broad pH optimum between pH 8.0 to 10.0. When the molarity of the tris(hydroxymethyl)aminomethane buffer was varied, the activity showed an optimum over the range 0.2 to 0.4 M. Activity was higher (135% of control level) when 20 to 80 mM Mg2+ was present. The Km for the enzyme was 0.25 mM. The D-alanine carboxypeptidase was inhibited by several beta-lactam antibiotics and the 50% inhibitory levels were 10(-8) M penicillin G, 10(-8) M ampicillin, 10(-5) M cloxacillin, and 5 x 10(-7) M methicillin.
Viral replication and interferon production in fetal and adult ovine leukocytes and spleen cells
Authors: Rinaldo C R , Overall J C , Glasgow L A
Published: January 2020

Peripheral blood leukocyte and spleen cell cultures derived from adult sheep and from third-trimester (107 to 145 days of gestation) and second-trimester (70 to 98 days of gestation) fetal lambs were examined for their ability to support viral replication and to produce interferon. Bluetongue virus, Herpesvirus hominis type 2, and Chikungunya virus failed to replicate in either leukocyte or spleen cell cultures derived from adult ewes or in cultures from second- or third-trimester fetal lambs. Similarly, peripheral blood leukocytes from adult sheep or third-trimester fetal lambs did not support the replication of Semliki Forest virus, vesicular stomatitis virus, Newcastle disease virus, or vaccinia virus. No major differences were observed in the ability of fetal and adult leukocytes to produce interferon in response to viral infection. In contrast, mean interferon titers induced by bluetongue virus, H. hominis type 2, and Chikungunya virus in spleen cells from second-trimester fetuses were 4- to 10-fold greater than those induced in spleen cells from adult ewes. Variations in interferon levels induced on separate occasions with cells from the same donor age group were observed. The antiviral substance induced in both the fetal and adult cell cultures fulfilled the usual criteria for characterization as interferon.
Comparative infectivity of trachoma organisms in HeLa 229 cells and egg cultures
Authors: Kuo C C , Wang S P , Grayston J T
Published: January 2020

Thirty-two trachoma strains were simultaneously titrated in the yolk sac of embryonated chicken eggs and in HeLa 229 cells pretreated with diethylaminoethyl-dextran to determine the relative sensitivity of the two culture systems for detection of the infection. The strains tested were both yolk sac- and HeLa cell-established isolates from the eye and the genital tract. The study showed that the cell culture was of equal or greater sensitivity than yolk sac culture and that ocular and genital isolates differ in their ability to propagate in egg and cell culture: genital strains grow better in cell culture than in eggs and ocular strains grow only slightly better in cell culture.
Intrauterine infection of mice with St. Louis encephalitis virus: immunological, physiological, neurological, and behavioral effects on progeny
Authors: Andersen A A , Hanson R P
Published: January 2020

Intravenous injection of pregnant mice with St. Louis encephalitis (SLE) virus at 8 days of gestation resulted in infection of the fetus. Progeny developed no antibody or tolerance to SLE virus since the viral antigen was cleared by maternal antibody before antibody-forming competence developed in the young. Temporary growth retardation was observed in a number of young at 3 weeks of age. After the initial setback the growth rate increased, indicating that early runting was due to an inability to adjust adequately to extrauterine life, which was subsequently overcome. In most other young there were no significant effects on growth, reproduction, or life expectancy. A few young died at or shortly after birth; in these, neurological changes ranging from gross defects such as encephaloceles and hydrocephalus to histological evidence of necrosis and congestion were observed. Neurologically related behavioral changes were detected by using the open field test and the rotating-rod test, which indicated neurological damage and memory impairment in the surviving intrauterinely infected animals.
Cytoplasmic antigens unique to the mycelial or yeast phase of Candida albicans
Authors: Syverson R E , Buckley H R , Campbell C C
Published: January 2020

Crossed immunoelectrophoresis with absorption in situ was used to distinguish the cytoplasmic antigens unique to the mycelial or yeast phase of Candida albicans from cytoplasmic antigens shared by both phases. The soluble cytoplasmic extracts of each growth phase had at least six distinct antigenic constituents not shared by the other phase. This technique is recommended for the analysis of closely related antigenic complexes.
Pathogenesis and immunology of experimental gonococcal infection: role of iron in virulence
Authors: Payne S M , Finkelstein R A
Published: January 2020

Addition of iron componds to inocula of the relatively avirulent T3 or T4 colony types of gonococci increased their lethality for chicken embryos after intravenous inoculation but had little or no effect on the highly virulent T1 or T2 types. The toxicity of nonviable inocula, killed cells or sonicates, was not significantly affected by ecogenous iron. Addition of the iron-binding protein conalbumin reduced or delayed the lethal effect of T1, but not T3, gonococci although growth of both colony types in the allantoic cavity of the embryo was inhibited by this protwin. This effect can be attributed specifically to deprivation of iron since the iron-complexed form of conalbumin had no apparent influence on growth or virulence. The results indicate that the ability to acquire iron in vivo is a significant factor in gonococcal virulence. The virulent colony types appear to have enhanced ability to compete with the host for iron and this may be related to the presence of pili, other surface components, or the synthesis of iron-chelating compounds.
Production and properties of an extracellular bacteriocin from Streptococcus mutans bacteriocidal for group A and other streptococci
Authors: Paul D , Slade H D
Published: January 2020

An extracellular bacteriocidal substance is produced by a serotype c strain of Streptococcus mutans in liquid meduim during the stationary phase of growth. The lethal effect of the substance was demonstrated by the decrease in viable counts of a standardized suspension of group A streptococci in broth. No lysis of affected cells was observed and no changes in appearance of these cells was seen in electron micrographs. The material was effective against certain strains of immmunological groups A, C, D, G, H, L, and O streptococci. It was inactive against strains of S. mutans belonging to the a, b, c, and d serotypes, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. The factor was purified 273-fold from the culture fluid by column chromatography. It was sensitive to trysin and Pronase and resistant to catalase. It possessed a molecular weight of more than 20,000 and was not dialyzable. The properties of this substance indicate that it is a bacteriocin. Group A streptococci, which had been treated with antiserum specific for the cell wall group and type antigens, were susceptible to the bacteriocin. Streptococcal strains resistant to the lethal action of the bacteriocin adsorbed the bacteriocin from the solutions, as did the sensitive cells. The bacteriocin was not adsorbed at 0 C.
Induction of herpes simplex virus immunity in newborn mice
Authors: Koyama H , Kasahara S
Published: January 2020

It was demonstrated that newborn mice, surviving an infection with attenuated herpes simplex virus, become resistant to challenge with a pathogenic strain of herpes simplex virus. This resistance did not seem to be mediated by antibody.
Cellular immunity in Peyer's patches of rats infected with Trichinella spiralis
Authors: Levin D M , Ottesen E A , Reynolds H Y , Kirkpatrick C H
Published: January 2020

A rat model of Trichinella spiralis gut infection was used to observe the sequence of developing cellular immunity in Peyer's patches and other lymphoid tissues. Whereas cellular reactivity (lymphocyte blastogenesis) for worm antigens was evident in mesenteric lymph nodes draining the gastrointestinal tract within 3 days after infection, Peyer's patch lymphocytes developed maximal reactivity 2 to 3 weeks later at the same time as the spleen and other lymphoid tissues. Furthermore, the immune reactivity found in Peyer's patches was only transient. Thus, in this parasitic gut infection, the Peyer's patch lymphoid tissue does not appear to be the first site of cellular responsiveness but rather to acquire cellular reactivity only when other lymphoid elements in the infected host have also acquired similar antigen-induced reactivity.
Ultrastructural observations on Ehrlichia equi organisms in equine granulocytes
Authors: Sells D M , Hildebrandt P K , Lewis G E , Nyindo M B , Ristic M
Published: January 2020

The ultrastructure of the etiological agent of equine ehrlichiosis, Ehrlichia equi, was studied in equine peripheral leukocytes. The organisms occurred within membrane-lined cytoplasmic vacuoles of neutrophils and eosinophils. Ovoid, round, and rod-shaped profiles were observed. From 1 to 33 organisms were present in a thin-section profile of a cytoplasmic vacuole. Many cells contained multiple organism-containing vacuoles. The organisms had a cell wall and plasma membrane, and internally they consisted of electron-dense and lucid areas. A great variation in size was observed. The morphological features were most consistent with agents of the genus Ehrlichia.
Effect of in vitro adaptation of Marek's disease virus on pock induction on the chorioallantoic membrane of embryonated chicken eggs
Authors: Sharma J M , Coulson B D , Young E
Published: January 2020

Cell-associated preparations of several isolates of Marek's disease virus produced more pocks on the chorioallantoic membrane of embryonated chicken eggs than plaques in duck embryo fibroblasts, thus indicating that lesion response in eggs was more sensitive than cytopathic response in duck embryo fibroblasts for assaying low-passage Marek's disease virus. Adaptation of the virus to cell cultures by serial passages, however, substantially reduced its pock response so that the titer ratio (plaque-forming units in duck embryo fibroblasts/pock-forming units in eggs) of cell culture-adapted Marek's disease virus was 1 or higher. The decreased pock response could not be attributed to selection of preexisting virus variant(s) with low affinity for chorioallantoic membrane because cloned Marek's disease virus had a good pock response at low cell culture passage levels, but this response decreased as the virus was attenuated by serial cell culture passage.
Comparison of antigens in sonic and pressure cell extracts of Mycobacterium tuberculosis
Authors: Janicki B W , Wright G L , Good R C , Chaparas S D
Published: January 2020

Comparisons were made of the yield, chemical content, and biological activity of filtrates and extracts obtained by sonic and pressure cell disruption of bacilli from 4- and 8-week-old Proskauer and Beck cultures of the H37Rv strain (TMC no. 102) of Mycobacterium tuberculosis. The culture filtrates were dialyzed, freeze-dried, reconstituted in saline, and sterilized by membrane filtration. The viable bacilli were washed and resuspended in distilled water and subsequently disrupted either by sonication in the cold for 15 or 30 min or by treatment at 20,000 or 40,000 lb/in2 in a pressure cell. The resulting extracts were clarified by centrifugation, concentrated, and sterilized by filtration. All preparations were adjusted to contain 10 mg of solids (dry weight)/ml and were analyzed quantitatively for protein, deoxyribonucleic acid, ribonucleic acid, polysaccharide, and lipid content. Separation patterns obtained by gradient acrylamide gel electrophoresis, as well as by one- and two-dimensional immunoelectrophoresis, provided the basis for qualitative comparisons of the culture filtrates and cell extracts. Three-point dose-response curves also were used to compare the preparations for skin test reactivity in BCG-vaccinated guinea pigs. It was concluded that, although there were no consistent differences in chemical content or biological activity between the preparations, a 15-min sonic treatment appeared to be the most suitable method for preparation of bacillary extracts based on yield of active components and ease of preparation.
Fate of interferon-treated cells
Authors: Friedman R M , Costa J R
Published: January 2020

Interferon-treated cultures of Ly cells survived initial infection with high multiplicities of vesicular stomatitis (VSV) or herpes simplex virus (HSV). In the case of HSV, infectious virus and intracellular viral antigen were rapidly eliminated from the interferon-treated cultures, and the cells grew out to form apparently normal monolayers that could be cultured indefinitely. In the VSV-infected Ly cultures, virus titers remained at low levels in interferon-treated cells but after about 14 days rapidly rose and the culture was destroyed. If interferon was added to the medium on days 4 and 6 after infection, virus titers rapidly declined but again recovered and the cells were destroyed. If, however, interferon treatment was resumed 9 days after initial infection, detectable infectious VSV was eliminated from the medium. Several methods, including cocultivation and molecular hybridization, failed to demonstrate persistence of a significant portion of the VSV genome in these cultures.
Distinct deoxyribonucleic acid polymerase activities associated with Dane particles and naked Dane cores
Authors: Bradley D W , Fields R M , Maynard J E
Published: January 2020

Two distinct deoxyribonucleic acid polymerase activities were found associated with liver fractions containing Dane particles and Dane cores.
Formation of cross-reacting antibodies against cellular and extracellular lipoteichoic acid of Streptococcus mutans BHT
Authors: Knox K W , Markham J L , Wicken A J
Published: January 2020

Antisera prepared against strains of Streptococcus mutans, particularly strain BHT, contain antibodies to the membrane lipoteichoic acid component that will cross-react with lipoteichoic acids from three species of lactobacilli. A more detailed study of the antibodies to strain BHT confirmed that the antibodies are specific for the polyglycerol phosphate component common to the lipoteichoic acids. Sera with a higher hemagglutinating titer could be obtained by injecting disrupted organisms. The partially purified lipoteichoic acid isolated from the culture fluid of S. mutans BHT was also immunogenic antibodies; antibodies were also formed against another unidentified component in this extracellular fraction.
Serum chemotactic inhibitory activity: heat activation of chemotactic inhibition
Authors: Epps D E , Williams R C
Published: January 2020

Serum chemotactic inhibitory activity (CIA) was studied in 46 patients with various systemic diseases, using a system consisting of normal human leukocytes as indicator cells and 10% fresh normal serum as a control chemotactic attractant. It was shown, as previously reported, that an association exists between CIA and skin test anergy. Heat treatment of sera at 56 C for 30 min increased both the incidence and the degree of chemotactic inhibition observed in these patients. The effects of heat treatment of sera containing CIA on other chemotactic attractants (C3a, bacteria-derived chemotactic factor (BF), and casein) are shown. Before heat treatment, some sera suppressed chemotaxis mediated by BF in the absence of suppression of normal serum-mediated chemotaxis, indicating the possible involvement of more than one system of inhibition. Multiple systems were further supported by data indicating that room temperature incubation resulted in a loss of CIA as measured by normal serum-mediated chemotoxis with no apparent decrease in the inhibition of BF -mediated chemotaxis. Separation of sera containing CIA by Sephadex G-200 showed chemotactic inhibitory activity to be increased in both the void volume region. Experiments showed that heat treating before separation resulted in similar increases in both peaks, implying the presence of an antagonist to CIA. Experiments demonstrating that sera containing CIA do not suppress casein-mediated chemotaxis by means of an irreversible inactivation of chemotactic factor are included along with experiments demonstrating a cellular mode of action. The possible presence of two systems of chemotactic inhibition, one acting directly upon chemotactic factors and one interacting with the responding cell, are discussed.
Bactericidal properties of human cell-free lymph
Authors: Martinez R J
Published: January 2020

The bactericidal properties of human cell-free lymph have been investigated. Gram-negative and gram-positive bacteria are susceptible to the killing action of lymph fluid. The colony-forming ability of Escherichia coli and Bacillus subtilis was reduced by 3 logs after 20 min of incubation with 10% (vol/vol) lymph. Salmonella and Listeria were less susceptible to killing. The presence of multiple killing factors in lymph showing different specificities for different bacterial cells was demonstrated.
Cytotoxic effect of the thermostable direct hemolysin produced by Vibrio parahaemolyticus on FL cells
Authors: Sakurai J , Honda T , Jinguji Y , Arita M , Miwatani T
Published: January 2020

The thermostable direct hemolysin produced by Vibrio parahaemolyticus showed cytotoxic activity on FL cells derived from human amniotic membrane. Scanning electron micrographs of the whole cells showed that the microvilli on the cell surface decreased in number and changed in shape on treatment with the hemolysin. Most of the microvilli disappeared before death of the cells, as judged from the results of staining the cells with trypan blue and measuring release of alkaline phosphatase from the cells. Electron micrographs of thin sections ofthe cells showed that the cytoplasm of the cells was not significantly affected by treatment with sublethal amounts of hemolysin, even when the microvilli on the cell surface were significantly affected. Lethal amounts of hemolysin affected the cytoplasm and caused disarovilli on the cell surface are affected by treatment with the hemolysin before cytoxic effects develop.
Pathological features of experimental gonococcal infection in mice and guinea pigs
Authors: Chandler F W , Kraus S J , Watts J C
Published: January 2020

The histopathological and immunofluorescent findings in tissues within and surrounding artificially created subcutaneous tissue cavities infected with Neisseria gonorrhoeae for 1 to 30 days were studied in mice and guinea pigs. Findings in the tissue cavities of the animal models were similar to the findings of disseminated gonococcal infection in humans. These similarities included an intense persistent polymorphonuclear leukocytic response with tissue necrosis, hemorrhage into the early lesion, a perivascular leukocytic response in adjacent tissue, difficulty in detecting large numbers of discrete morphologically typical gonococci by the tissue Gram stain and direct fluorescent antibody techniques, a decrease in the number of identifiable gonococci with duration of the infection, and moderate amounts of extracellular and intracellular immunofluorescent gonococcal debris. Studies into the pathogenesis of the animal infections may enhance our understanding of the pathogenic mechanism (s) associated with gonococcal infection in humans.
Growth and effects of ureaplasmas (T mycoplasmas) in bovine oviductal organ cultures
Authors: Stalheim O H , Proctor S J , Gallagher J E
Published: January 2020

Ureaplasmas isolated from the human genital tract and from the genital and respiratory tracts of cattle were grown in association with organ cultures of bovine oviduct (uterine tube). All strains of unreaplasmas multiplied in organ cultures, stopped ciliary activity, and caused histological lesions. Most strains grew well, and 10(8) to 10(9) color-changing units were determined 18 to 144 h after inoculation. Twenty-four to 144 h after inoculation with unreaplasmas, ciliostasis was complete. Ciliostasis was also caused by additions of nonviable cultures at pH 8.8 (or adjusted to 7.4) or washed disrupted cells (100 mug of protein/ml); it occurred in 48 to 96 h. The cilia-stopping effect of nonviable cultures was diminished by heating (56 C for 30 min) and was abolished by boiling. When added to fresh medium in amounts exceeding 25%, nonviable unreaplasmal cultures completely inhibited ureaplasmal growth. By light, scanning, and transmission electron microscopy, cilia-stopping effect was correlated with collapse and sloughing of the cilia (the initial lesion was "bent" cilia), with bulging and vacuolization of secretory and ciliated cells, and finally with disorganization of the epithelium, necrosis, and desquamation.
Evaluation of adenosine deaminase and other purine salvage pathway enzymes in horses with combined immunodeficiency
Authors: McGuire T C , Pollara B , Moore J J , Poppie M J
Published: January 2020

Foals with combined immunodeficiency had normal levels of purine salvage pathway enzymes, including adenosine deaminase, nucleoside phosphorylase, and xanthine oxidase.
Some effects of silical treatment on Marek's disease
Authors: Higgins D A , Calnek B W
Published: January 2020

Treatment of newly hatched chicks with silica by the intraperitoneal route delayed the onset of mortalities due to the JM strain of Marek's disease (MD' virus inoculated at 6 days of age. During the 88-day observation period fewer silica-treated chicks died of MD, but this difference was not usually statistically significant. Silica treatment had no effect on the susceptibility of 4-week-old birds. Silica treatment reduced the antibody response to MD but, in general, not significantly. The antibody response to bovine serum albumin was significantly enhanced if measured by the indirect hemagglutination test but not if measured by the agar gel diffusion test, whereas the response to Brucella abortus was enhanced significantly in N-line (MD-resistant) chicks but not significantly in P-line (MD-susceptible) chicks. Five days after infection, silica-treated chicks had significantly less fluorescing antigen in thymus and bursa than did untreated chicks; no difference was observed in the spleen. After silica treatment the glass-adherent cell population in the buffy coat was increased by up to 10-fold compared with untreated chicks. It is suggested that silica treatment induced macrophage proliferation, with subsequent restriction of MD virus spread, yet allowed an adjuvant-type effect with other antigens.
Diethylaminoethyl-cellulose-bacterial cell immunoadsorbent columns: preparation of serotype-specific globulin and immunofluorescent conjugates for Streptococcus mutans serotypes a and d
Authors: McKinney R M , Thacker L
Published: January 2020

Diethylaminoethyl (DEAE)-cellulose was used as a support material for preparing bacterial cell columns. Pretreatment of the bacterial cells with formalin was essential in obtaining satisfactory adherence of the cells to DEAE-cellulose. Cross-reacting antibodies were removed from antibody preparations against strains of Streptococcus mutans serotypes a and d by adsorption on appropriate bacterial cell columns. S. mutans serotype d was further divided into two subtypes on the basis of immunofluorescent staining with conjugates of immunospecifically adsorbed immunoglobulin G. The DEAE-cellulose-bacterial cell columns were regenerated after use by desorbing the cross-reacting antibodies with low-pH buffer and were used repeatedly over and 18-month period with no detectable loss in effectiveness.
Major outer membrane protein of Campylobacter fetus: physical and immunological characterization
Authors: McCoy E C , Wiltberger H A , Winter J
Published: January 2020

The outer membrane proteins of Campylobacter fetus have been isolated by extraction of cell envelopes both in Triton X-100 and ethylenediaminetetraacetate (EDTA) and in sodium dodecyl sulfate (SDS). Each method yielded a major protein component, which migrated identically in SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and proved comparable in electrophoretic and molecular weight characteristics to the analagous protein from Escherichia coli. In addition, the surface of SDS-extracted C. fetus cells displayed a subunit structure similar to that observed in E. coli. The major envelope protein isolated with SDS appeared antigenically identical with one of the proteins isolated with Trition-EDTA on the basis of immunodiffusion reactions with specific antisera. Antibodies directed to the major envelope protein were not reactive in agglutination, immobilization, bactericidal, or opsonization reactions. Strains of C. fetus belonging to each of the three O serotypes possessed major envelope proteins comparable in SDS-PAGE but distinguished antigencally in a fashion paralleling the O serotype classification.
Acquired immunity to Aspergillus fumigatus
Authors: Lehmann P F , White L O
Published: January 2020

In mice preinfected with Aspergillus fumigatus for greater than or equal to days, administration of cortisone failed to stimulate mycelial growth. The liver and heart of mice preinfected for 14 days resisted infection after a cortisone + conidia challenge.
Detection of the K99 antigen by means of agglutination and immunoelectrophoresis in Escherichia coli isolates from calves and its correlation with entertoxigenicity
Authors: Guinée P A , Jansen W H , Agterberg C M
Published: January 2020

The common antigen of calf enterotoxigenic strains of Escherichia coli, recently established as the K99 antigen, was studied by means of the slide agglutination test and immunoelectrophoresis. Specific antisera were obtained by absorption of crude antisera with ultrasonicates of autologous cells grown at 18C or by injection into rabbits of the purifies K99 antigen obtained by preparative electrophoresis. The K99 antigen was usually undetectable in calf enterotoxigenic E. coli cultures with capsular K antigens of the A variety grown at 37C on commercially available nutrient agar plates designed for the isolation of Enterobacteriaceae, but was rapidly detectable when grown on a buffered semi-synthetic medium at pH 7.5 (Minca medium). An alternative procedure for the isolation and identification of calf entertosigenic E. coli strains from feces, using the Minca medium, is proposed. K99 was found in 70 of 74 strains of E. coli, the enterotixigenicity of which was established in the ligated gut test in calves. None of the 20 cultures negative in the ligated gut test possessed K99 antigen. The K99 antigen is therefore probably a useful diagnostic tool for the identification of calf enterotixigenic E. coli strains, taking into account that K99 and enterotoxigenicity are controlled by different plasmids.
Modification of resistance of mice to Naegleria fowleri infections
Authors: Adams A C , John D T , Bradley S G
Published: January 2020

Naegleria fowleri, which produces a fatal meningoencephalitis in humans, is also able to produce a progressive and fatal disease in mice. The course of the disease in DUB/ICR mice is dependent upon the infecting dose of organisms, whether administered intraperitoneally (i.p.) or intravenously (i.v.). All of the mice receiving 10(7) trophozoites/mouse i.v. or 4.85 X 10(7) trophozoites/mouse i.p. were killed within 10 days. Escherichia coli O26:B6 lipopolysaccharide, administered at a dose of 1 mg/kg 24 h prior to N. fowleri, afforded some protection for several days after challenge, but by day 8 there was no difference in survival of untreated and endotoxin-treated mice. No significant protection was afforded by a complex of lipid A with concanavalin A (ConA) or bovine serum albumin (BSA) or by dimethylmyristamide-BSA, dimethylmyristamide, BSA, beta-hydroxymyristic acid-ConA, beta-hydroxymyristic acid, ConA, myristic acid-BSA, or myristic acid. Mice surviving primary i.v. or i.p. challenge doses of N. fowleri, 5 X 10(6) and 10(7) trophozoites/mouse, respectively, were highly resistant to rechallenge with an i.v. dose of organisms (5 X 10(6) Naegleria/mouse) that produced uniformly fatal disease in untreated control mice.
Interaction of diphtheria toxin and its active subunit, fragment A, with toxin-sensitive and toxin-resistant cells
Authors: Moehring T J , Moehring J M
Published: January 2020

Diphtheria toxin and purified fragment A, the active subunit of the toxin, were tested on toxin-sensitive and permeability class toxin-resistant cultured mammalian cells. Protein synthesis was inhibited to the same degree in sensitive or resistant cells by active concentrations of purified fragment A. In contrast, resistant cells required a concentration of whole toxin 5 to 6 logs greater than that required by sensitive cells to achieve the same degree of inhibition. On a molar basis, the toxicity of fragment A was equivalent to that of whole toxin on resistant cells. These results are evidence for the existence of two independent mechanisms for the entry of toxin or its active moiety into cells. One mechanism is a highly efficient, toxin-specific entry mechanism, involving surface receptor and fragment B-mediated association, and is active in sensitive cells. The other is a less efficient, nonspecific mechanism, probably related to endocytosis, which is operative in sensitive and resistant cells but is inapparent in sensitive cells when they are exposed to whole toxin because of the action of the specific mechanism.
Use of the hemadsorption phenomenon for determining virus and neutralizing antibody titers of rabies
Authors: Minamoto N , Kurata K , Kaizuka I , Sazawa H
Published: January 2020

Chicken embryo cells infected with the HEP Flury strain of rabies virus adapted to tissue culture produced a hemadsorption (HAD) phenomenon by using goose erthyrocytes. The optimal conditions for HAD included the incubation of cell cultures at 37C for 3 days after virus inoculation, the use of a 0.4% suspension of goose erythrocytes in phosphate buffer adjusted at pH 6.2, and adsorption of erythrocytes at 4C. This phenomenon was inhibited with anti-rabies serum. Virus titer obtained with the HAD technique was almost the same as with the fluorescent antibody technique or the intracerebral inoculation of suckling mice. Results of the neutralization test by using the HAD technique could be easily determined 3 days after inoculation of chicken embryo cells with the mixture of 100 mean tissue culture infective doses of virus and diluted serum. The neutralizing antibody titers coincided with those obtained in mice.
Cell-mediated immunity to varicella-zoster virus demonstrated by viral inactivation with human leukocytes
Authors: Gershon A A , Steinberg S , Smith M
Published: January 2020

Cell-mediated immunity to varicella-zoster (V-Z) virus in persons immune to varicella has been demonstrated, using a tissue culture technique. Cell-mediated immunity was reflected by the ability of peripheral leukocytes (lymphocytes and monocytes) from human donors to inactivate V-Z virus. Leukocytes were stimulated by the addition of noninfectious V-Z antigen to cultures newly infected with V-Z virus. Several days leter, the V-Z virus in these cultures was titered. When leukocytes from donors immune to caricella were used, a significant decrease in V-Z titer, compared with controls, was noted. When leukocytes from donors susceptible to varicella were tested, no decrease in V-Z virus titer was found. A mixed population of lymphocytes and monocytes from immune donors was required to demonstrate inactivation of V-Z virus. The development of specific cell-mediated immunity to V-Z virus may play a role in termination of varicella and in prevention of second attacks of this disease.
Role of platelets in the pathogenesis of canine endotoxin shock
Authors: From A H , Fong J S , Chiu T , Good R A
Published: January 2020

Endotoxin-platelet interactions are thought to be of major importance in the response of dogs and other species to bacterial endotoxin; the mechanisms postulated are: (i) the release of vasoactive substances, (ii) the formation of occlusive platelet aggregates, and (iii) induction of intravascular coagulation. The role of platelets in canine endotoxin shock was examined in animals with thrombocytopenia induced by estrogen pretreatment (less than 10,000 platelets/mm3) and in controls. After intravenously administered endotoxin, the hemodynamic responses, mortality, and gross necropsy findings were similar in both groups. These data indicate that endotoxin-platelet interactions are not determinative in the pathogenesis of canine endotoxin shock.
Fate of 51Cr-labeled lipopolysaccharide in tissue culture cells and livers of normal mice
Authors: Zlydaszyk J C , Moon R J
Published: January 2020

Livers of normal mice trapped over 80% of intravenously injected 51Cr-labeled lipopolysaccharide after 1 h. Liver fractionation studies showed that nearly 45% of the labeled endotoxin was associated with cell nuclei, 20% with the mitochondrial-lysosomal fraction, and approximately 30% with the cell sap. Analysis of the distribution of 51Cr-labeled lipopolysaccharide among parenchymal and Kupffer cells showed that over 75% of the in vivo-trapped counts were parenchymal cell associated. Cell populations were approximately 65% parenchymal cells and 35% nonparenchymal cells. Further, six non-reticuloendothelial system tissue culture cell lines were tested for their ability to internalize labeled lipopolysaccharide. In all cells studied, 1 to 4% of the labeled lipopolysaccharide was taken up after 3 h, with greater than 80% of the counts localized in the nuclear fraction. The data show that non-reticuloendothelial system cells can sequester endotoxin both in vivo and in vitro and suggest that parenchymal cells as well as Kupffer cells remove circulating endotoxin from the blood.
Interactions of Pseudomonas aeruginosa with immunoglobulins and complement in sputum
Authors: Hann S , Holsclaw D S
Published: January 2020

The interactions of Pseudomonas aeruginosa with humoral factors in the sputum of patients with cystic fibrosis were investigated by using an indirect immunofluorescent technique. Fluorescein-conjugated, monovalent antiserum specific to heavy chains of human immunoglobulin A (IgA), IgG, or IgM and to complement C3 were used. All strains of P. aeruginosa recovered from the sputum specimens of patients with cystic fibrosis were found to be coated with antibodies of IgA, IgG, and IgM classes and with C3. The specificity of the antibody coating was determined. The fluorescence was most intense with IgA and was followed in intensity by IgG, IgM, and C3. No difference was noted between rough and mucoid strains of P. aeruginosa. When the subcultured P. aeruginosa was incubated with the sputum eluates, a similar pattern of fluorescence was demonstrated, indicating that these humoral factors are present in the sputum and that the coating process can take place in the lower respiratory tract of the patients. By single radial immunodiffusion, significant quantities of the humoral factors in the sputum eluates were detected. These findings suggest that P. aeruginosa is opsonized in sputum of patients with cystic fibrosis.
Granulocyte function and Pseudomonas burn wound infection
Authors: McEuen D D , Gerber G C , Blair P , Eurenius K
Published: January 2020

Granulocyte function in burn injury has been evaluated with an in vitro animal assay. Bactericidal activity, using this technique, was markedly depressed only when granulocytes from scald burned rats were challenged with each of three strains of Pseudomonas aeruginosa in the presence of autologous postburn serum or heat-inactivated serum. Preincubation of postburn and normal serum at 37 degrees C for 1 h resulted in normal phagocytosis. These studies validate an investigation of plasma and/or granulocyte replacement therapy in the burned patient.