bluuebunny/crossref_metadata_2025 huggingface dataset:
Passive immunity against pseudomonas sepsis during granulocytopenia
Authors: Harvath L , Andersen B R , Amirault H J
Published: January 2020

Specific passive immunity against Pseudomonas aeruginosa sepsis was assessed in granulocytopenic dogs. Dogs were infused with either normal or antipseudomonas immune plasma 24 h before pseudomonas challenge. They were challenged intravenously with 10(7) serotype 6 P. aeruginosa during granulocytopenia. Treatment was evaluated by observation of survival periods, febrile responses, type 6 pseudomonas antibody titers, and quantitative cultures of blood and tissues. The results demonstrated that passively immunized dogs did not survive infection. Both normal-plasma and immune-plasma recipients had bacteremia at death, with median values of 980 and 470 pseudomonas per ml of blood, respectively. All dogs had marked febrile responses 24 h after pseudomonas challenge and had high concentrations of pseudomonas in their lung tissue at death, with median values of 10(8) pseudomonas per g of wet tissue weight. After plasma infusion, immune-plasma recipients had high concentrations of anti-pseudomonas antibody, with total antibody titers ranging from 256 to 1,024 and a median value of 1,024. These titers were comparable to titers attained in a previous study from our laboratory using active immunization with pseudomonas lipopolysaccharide vaccine, where the median total anti-pseudomonas antibody titer was 2,048. Actively immunized animals, however, were significantly protected against pseudomonas sepsis and had prolonged survival periods and prevention of bacteremia. The present study demonstrates that circulating type-specific antibody is not solely responsible for the protection afforded to granulocytopenic dogs actively immunized against pseudomonas.
Limulus amebocyte lysate reaction with streptococcal pyrogenic exotoxin
Authors: Brunson K W , Watson D W
Published: January 2020

Three immunologically distinct pyrogenic exotoxins derived from group A streptococci were found to cause coagulation of limulus amebocyte lysate. These exotoxins were shown to be several thousand times less active for lysate coagulation than gram-negative endotoxin.
Chromosomal determinants for exfoliative toxin production in two strains of staphylococci
Authors: Rosenblum E D , Tyrone S
Published: January 2020

Strain TG is a plasmidless exfoliative toxin (ET) producer. Strain ER201 contains a plasmid, whose loss had no apparent effect on ET production. The plasmid of ER201 was almost identical in size to the plasmid from strain UT0007, which carries an ET determinant.
Microbicidal mechanisms of human granulocytes: synergistic effects of granulocyte elastase and myeloperoxidase or chymotrypsin-like cationic protein
Authors: Odeberg H , Olsson I
Published: January 2020

The antibacterial activity of a myeloperoxidase (MPO)-glucose oxidase system was found to be greatly increased by granulocyte elastase, present in azurophil granules of human neutrophils. The MPO-H2O3-mediated killing of both Escherichia coli and Staphylococcus aureus was potentiated by granuocyte elastase at an acid pH, whereas at pH 7.4 only killing of E. coli was potentiated. The potentiating effect of elastase was not dependent on the enzymatic properties of the protein since it was not abolished by heating, which destroys the enzymatic activity. A peptide chloromethyl ketone elastase inhibitor abolished both elastolytic activity and the pctentiating effects on MPO-H2-O2-mediated bacterial killing. The antibacterial activity of chymotrypsin-like cationic protein of human neutrophils was also potentiated by elastase. Other degradative enzymes isolated from human granulocytes, e.g., collagenase and lysozyme, did not potentiate MPO-H2O2-mediated or cationic protein-dependent bacterial killing. The present study indicates that a neutrophil constitutent, elastase, which is not microbicidal by itself, can initiate sublethal changes that render some microorganisms more susceptible to the action of microbicidal agents like MPO and chymotrypsin-like cationic protein.
Hepatic clearance of Candida albicans in rats
Authors: Sawyer R T , Moon R J , Beneke E S
Published: January 2020

The initial clearance of Candida albicans from the blood stream of rats and from perfusion medium by perfused rat livers was characterized. Normal rats cleared over 90% of large doses of intravenously injected yeast cells in 5 min. All were recovered as viable cells among various reticulendothelial organs after 30 min. The perfused rat liver trapped an average of 85% of the yeast cells in a single pass. No significant killing occurred, even in the presence of 10% whole rat blood. Scanning electron microscopy of cryofractured livers revealed that the cells were trapped in liver sinusoids but outside phagocytic cells.
Depressed antibody responses to a thymus-dependent antigen in toxoplasmosis
Authors: Strickland G T , Sayles P C
Published: January 2020

The immunodepressive effect of Toxoplasma gondii infection in mice was studied, using sheep erythrocytes (SRBC) as the testing antigen and serum hemagglutinins, hemolysins, and both direct and indirect splenic plaque-forming cells (PFC) to SRBC as assays. In the primary antibody response, immunoglobulin M (IgM), hemagglutinins, and hemolysins and both IgM- and IgG-secreting PFC were depressed in animals immunized after infection. Maximum immunodepression occurred during the first 3 weeks of Toxoplasma infection. When the secondary antibody response was studied, results varied. Mice immunized with SRBC after being infected with T. gondii had a depression in both IgM and IgG PFC. Mice immunized with SRBC before being infected with T. gondii and then given a challenge dose of SRBC had a delay, but no an actual depression, in IgG hemagglutinins and hemolysins and IgG-secreting PFC. These studies show that the immunodepression associated with Toxoplasma infection is complicated, and they provide no definitive explanation for the mechanism.
Stable L-Forms of
Clostridium perfringens
: Growth, Toxin Production, and Pathogenicity
Authors: Mahony D. E.
Published: January 2020

Growth and toxin production of stable L-forms of
Clostridium perfringens
grown in a mini-fermentor were monitored. A gradual but steady increment in viable count occurred over a 7-h period, followed by death. The peak of viability preceded the optical density peak by 3 h. Theta, alpha, kappa, and lambda toxins were measured, with theta toxin appearing first in the culture supernate. Growth of the parent bacillus form of
C. perfringens
was compared under similar conditions. Toxin levels achieved by the bacillus culture exceeded those of the L-form culture four- to eightfold; however, based upon viable count, the L-form organism produced 8 to 16 times as much toxin as did the bacillus. The amounts of extracellular toxin produced by both forms were similar when related to cell protein rather than cell number. Guinea pig inoculation showed that the L-form of
C. perfringens
did not produce gas gangrene, although it was not entirely without effect. Both guinea pig and human sera were inhibitory to these L-forms, a fact attributable to a heat-liable component in the sera, most likely complement.
Expression of Aleutian mink disease antigen in cell culture
Authors: Hahn E C , Ramos L , Kenyon A J
Published: January 2020

Infection of CRFK feline kidney cells with Aleutian disease vurus leads to production of virus-induced antigen(s) in the nucleus which could be demonstrated by the fluorescent-antibody technique. The number of fluorescent nuclei was lineraly dependent on the dilution of the inoculum, but rarely exceeded 20% of the cells. Aleutian disease nuclear antigen was only transiently detectable. The virus-induced antigen was detected after infection of cells of several divergent species; however, the CRFK line of feline kidney cells was the most susceptible. Inhibitor studies indicated that deoxyribonucleic acid synthesis, ribonucleic acid synthesis, and protein synthesis were required for viral antigen production. Cell growth was also a requirement for synthesis of viral antigen, An in situ radioimmune assay was used to measure binding of 125I-labeled mink anti-Aleutian disease virus to infected cells and competition with unlabeled sera. The system is suitable for quantitation of infectivity.
Hemagglutinating activity of Fusobacterium nucleatum
Authors: Falkler W A , Hawley C E
Published: January 2020

Gingival isolates of oral Fusobacterium nucleatum strains (gram-negative anaerobic fusiform bacilli) have shown the characteristic ability to hemagglutinate a variety of erythrocytes (RBC) of human and animal origin. Other members of the genus tested (F. necrophorus, F. varium, and F. mortiferum) displayed little if any ability to hemagglutinate RBC. The hemagglutination (HA) activity could be observed in the F. nucleatum strains with the whole cells and in most instances with sonicated preparations of the organisms. The HA activity was observed in cell wall preparations of the organism and appeared dependent upon a heat-labile protein component of the cell wall. In decreasing order, the RBC that would hemagglutinate with the smallest concentration of HA preparations were rabbit, monkey, human, sheep, horse, and ox. No differences in HA activity of the preparations with cells from the various human blood types were noted. Absorption of the HA preparation of one strain with human cells removed HA moiety was bound to the cells via a Ca2+ binding site interaction since ethylenediaminetetraacetic acid and ethylene glycol-bis-N,N'-tetraacetic acid inhibited binding, and HA could be reestablished by the addition of Ca2+ but not Mg2+. Rabbit antisera to the F. nucleatum strains inhibited HA activity when tested with the HA preparation in the standard test, whereas anti-Leptotrichia buccalis sera or normal rabbit sera had no effect. A tanned-cell passive HA test with rabbit anti-F. nucleatum sera displayed reactivity between the homologous strains but little reactivity with the other Fusobacterium species tested.
Congenital abnormalities in newborn lambs after infection of pregnant sheep with Akabane virus
Authors: Parsonson I M , Della-Porta A J , Snowdon W A
Published: January 2020

Akabane virus (a Bunyavirus) has been associated with epizootics of congenital deformities in cattle, sheep, and goats. Experimental studies using mouse-adapted virus inoculated intravenously into pregnant sheep gave an inapparent infection. Neutralizing antibodies were detected on day 5, and peaks in the titer were seen at days 10 and 48. Ewes infected at day 30 to 36 of pregnancy produced five (31% incidence) deformed lambs. Sera from four of these possessed neutralizing antibodies to Akabane virus before ingesting colostrum. Two lambs had arthrogryposis, hydranencephaly, kyphosis, scoliosis, and brachygnathia; one had micrencephaly; and the other two had porencephaly. The two lambs with arthrogryposis and hydranencephaly also had extensive lesions in other tissues. In the spinal cord there was a marked decrease in the number of ventral horn neurones and a depletion of myelin. Skeletal muscles showed marked atrophy. The medulla of the thymus possessed large Hassall's corpuscles and a reduced number of thymocytes in the cortex. It would appear that the pathogenic effects of Akabane virus are related to the gestational age (30 to 36 days) at which the fetus is infected. Akabane virus can now be included in the growing list of teratogenic viruses and provides an interesting system for studying such congenital diseases.
Mitogenic stimulation of murine spleen cells: relation to susceptibility to Salmonella infection
Authors: von Jeney N , Günther E , Jann K
Published: January 2020

The screening of several inbred strains of mice suggested that the capacity of their spleen cells to respond to the mitogenic effect of lipopolysaccharide (LPS) of gram-negative bacteria was correlated with their resistance to intraperitoneal infection with Salmonella typhimurium. An infection of LPS into mice caused changes in the in vitro responsiveness of their spleen cells to the mitogenic effects of LPS and phytohemagglutinin. Pretreatment of mice with whole ultraviolet (UV)-killed bacteria led to a marked rise in the in vitro response of the spleen cells to UV-killed bacteria, but not to LPS or or phytohemagglutinin. This enhanced response to UV-killed bacteria was not specific for the O antigens of the bacteria.
Cell walls of Coccidioides immitis: neutral sugars of aqueous alkaline extract polymers
Authors: Wheat R , Scheer E
Published: January 2020

The neutral sugar components of hydrolysates of the alkali-soluble and -insoluble fractions of an acid-extracted Coccidioides immitis mycelial cell wall preparation were examined by gas-liquid chromatography. The alkali-soluble fraction was separated into a neutral water-soluble (46% carbohydrate) and a neutral water-insoluble fraction (6% carbohydrate). Glucose was the major neutral sugar constituent of all fractions. Mannose appeared to be a second major component of the alkali-soluble, water-soluble fraction. 3-O-methylmannose was absent. Small amounts of other sugars, including galactose and pentoses, were tentatively identified. Borohydride reduction before alkaline extraction resulted in retention of almost half of the glucose and variable amounts of other components in the alkaline residue, indicating that solubilization of cell wall polymers by alkaline extraction results in significant degradation of the native cell wall polymers.
Production and detection of staphylococcal elastase
Authors: Hartman D P , Murphy R A
Published: January 2020

The optimum conditions were determined for the production and detection of staphylococcal elastase from Staphylococcus epidermidis. Optimum production and recovery took place when the dialysis membrane technique was utilized with brain heart infusion agar incubated for 44 h under 15% CO2 and harvested in physiological saline. Near-optimal production took place by 28 h, and this was found more useful for routine use. Incorporation of elastin into the culture medium or the inoculum did not result in higher levels of elastase production. The detection system consisted of 0.25% particulate elastin suspended in a pH 7.0, 0.05 M tris(hydroxymethyl)aminomethane-hydrochloride-buffered solidified plated medium. Crude undialyzed elastase was best detected when the medium contained either agarose and 10(-3) M calcium or purified agar without additional additives. Crude dialyzed elastase was best detected when the medium contained agarose and 10(-3) M disodium ethylenediaminetetraacetic acid.
Altered immune responsiveness associated with Encephalitozoon cuniculi infection in rabbits
Authors: Cox J C
Published: January 2020

The variation in immune response of two unrelated colonies of laboratory rabbits to high doses of heat-killed Brucella abortus strain 19 was investigated. One was a mixed-breed, multicolored colony in which a high prevalence of encephalitozoonosis had been recorded, whereas the other rabbits were derived from a colony of Dutch-marked specific-pathogen-free rabits. Although considerable variation in the immune response between individual rabbits was noticed at all bleeds, rabbits infected with Encephalitozoon cuniculi showed, on comparison with uninfected rabbits from either colony, a depressed immunoglobulin G response from week 5 of the antigen injection schedule and, from week 8, an elevated immunoglobulin M response.
Purification of a monocytosis-producing activity from Listeria monocytogenes
Authors: Galsworthy S B , Gurofsky S M , Murray R G
Published: January 2020

The monocytosis-producing activity (MPA) from Listeria monocytogenes is shown to be associated with the cell envelope. Both MPA and immunosuppressive activity were readily extracted with aqueous solvents and separated as two independent activities by gel filtration and ion-exchange chromatography. As little as 10 microng of the resulting MPA-containing fraction caused a ninefold elevation in the level of circulating monocytes. The molecular weight of the fraction was approximately 22,000, and it contained phosphorus and carbohydrate.
Use of fluorescent antibody in studies of immunity to cholera in infant mice
Authors: Guentzel M N , Field L H , Eubanks E R , Berry L J
Published: January 2020

Infant mice 8 days of age were infected orally with virulent, motile, classical or El Tor strains of Vibrio cholerae and with nonmotile mutants of low virulence derived from the same strains. At intervals of 8 and 12 h postinfection, frozen thin sections of the ileum were prepared, stained with fluorescein isothiocyanate-labeled rabbit anti-vibrio antibody, and examined with the fluorescence microscope. The motile organisms were present in larger numbers, especially at 12h, and had penetrated the intervillous spaces and crypts of Lieberkuhn more completely than nonmotile vibrios. Dilution counts were made on various regions of the intestines of infant mice challenged orally 12 h previously with either motile or nonmotile strains of V. cholerae. Greater numbers of organisms were found, especially in the upper intestinal regions, when motile organisms were used. Low numbers of vibrios, limited mostly to the lumen, were seen in the ileum of infant mice infected with motile organisms when the infants were the offspring of mothers that had been immunized with crude flagellar vaccine or a vesicular preparation derived from the vibrio cell surface. The distribution of vibrios in this case was similar to that found in infected infants of unvaccinated mothers challenged with nonmotile organisms. Motility appears to enable the bacteria to better populate the upper regions of the intestinal tract and to avoid the washing effects of secretions and peristalsis. Antibacterial immunity may function, at least in part, by making it impossible for motile vibrios to accomplish this widespread distribution within the ileum.
Relationship between Habs serotypes and 2-amino sugar composition of Pseudomonas aeruginosa
Authors: Suzuki N , Tsunematsu Y , Matsumoto H , Tazaki T
Published: January 2020

Thirteen Habs-serotype strains could be classified into 11 groups depending on the characteristic patterns of 2-amino sugar composition; strains of serotypes 2, 5, and 7 had the same pattern, and each other serotype strain had its own distinctive pattern. In our classification, fucosamine and quinovosamine were of importance. We found that fucosamine from strain P14 (Habs serotype 1) was of the D configuration only, unlike DL-fucosamine from strain N10 (Habs serotype 11).
Effect of water temperature on a herpesvirus infection of sea turtles
Authors: Haines H , Kleese W C
Published: January 2020

The role of water temperature in the induction and maintenance of a dermal herpesvirus infection (gray-patch disease) of young, green sea turtles (Chelonia mydas) was studied under carefully controlled experimental conditions, in which the influence of other recognized stress factors was negligible. A nimals that were subjected to a gradual temperature increase from 25 to 30 degrees C, with subsequent maintenance at 30 degrees C, and those that were abruptly shifted from water at 25 degrees C to water at 30 degrees C showed a significantly shorter period before the onset of clinical signs and an increase in the severity of the lesions when compared with control animals. Animals that were subjected to a gradual increase in water temperature from 25 to 30 degrees C and a subsequent decrease to 25 degrees C, where they were maintained, had a period before onset of clinical signs and severity closer to that of control animals. Our findings indicate that both the induction of clinical gray-patch disease and the severity of the lesions are affected by water temperature and suggest that one possible means of control of this herpesvirus infection under intensive aquaculture conditions might be water temperature manipulation.
Antigenic relationships among several simian varicella-like viruses and varicella-zoster virus
Authors: Felsenfeld A D , Schmidt N J
Published: January 2020

Cross-neutralization and complement fixation tests demonstrated the immunological identity of the Delta herpesvirus, the 592S virus, the Liverpool vervet monkey virus, the herpesvirus of patas monkeys, and the Medical Lake macaque virus. These viruses were isolated from diverse outbreaks of varicella-like disease in simians and from various simian species. All of the simian viruses were shown to be related to human varicella-zoster (V-Z) virus, as evidenced by the fact that immunization of monkeys with each of the simian viruses elicited the production of both neutralizing and complement-fixing antibodies to V-Z virus. However, cross-complement fixation tests indicated that the simian viruses are not so closely related to V-Z virus as they are to one another. Varicella or zoster infections in humans produced neutralizing and complement-fixing antibody responses to each of the simian viruses; the responses were more marked in zoster infections than in varicella infections but, in most patients, antibody levels produced to the simian viruses were much lower than those to the homologous V-Z virus.
Ethylenediaminetetraacetic acid-sensitive antiphagocytic activity of Neisseria gonorrhoeae
Authors: Rosenthal R S , Fulbright R S , Eads M E , Sawyer W D
Published: January 2020

Colonial types of Neisseria gonorrhoeae were examined for the presence of pilus-independent antiphagocytic activity. Type 3 and depiliated type 1 gonococci had a shearing- and protease-resistant antiphagocytic activity that was eliminated by treatment with ethylenediaminetetraacetic acid (EDTA) and that was not present on type 4 bacteria. Incubation of EDTA-treated bacteria 37 degrees C for 90 min resulted in fas prevented by antibiotics that block the final assembly of cell wall macromolecules that depend on the C55-isoprenoid carrier for export. These include both lipopolysaccharide and peptidoglycan. Restoration was, however, unaffected by drugs that interfere with the synthesis of peptidoglycan, but not that of lipopolysaccharide, and by inhibitors of protein synthesis. These data suggested that gonococci have an antiphagocytic mechanism in addition to the previously described determinant (presumably pili) that was removed by blending or by treatment with proteases. Of the two antiphagocytic activities, type 1 had both, type 3 had only the EDTA-sensitive component, and type 4 had neither.
In vitro response of rabbit alveolar macrophages to infection with Nocardia asteroides
Authors: Beaman B L
Published: January 2020

The interaction of Nocardia asteroides with cultured "normal" nonimmune rabbit alveolar macrophages was studied by light and electron microscopy. It was shown that the alveolar macrophage response to the more virulent strain (N. asteroides 14759) was quite different from the response to the less virulent organism (N. asteroides 10905). N. asteroides 14759 elicited a dramatic in vitro response of the macrophages toward the nocardial infection. Within a few hours postinfection, there was a migration of macrophages toward other cells actively infected with viable nocardia, so that at 6 h considerable macrophage aggregation on the cover slips had occurred. Many of the macrophages within these aggregates exhibited tight cell-to-cell contact, whereas others were observed to fuse, forming multinucleate giant cells, with many containing more than 10 nuclei. Upon continued incubation, these giant cells appeared to destroy the intracellular nocardia, so that, at 24 h postinfection, gram-positive, ultrastructurally intact bacteria could not be observed. At the same time, some of the macrophage aggregates that did not fuse appeared to be unable to stop the intracellular growth of nocardia. At 12 to 24 h large numbers of gram-positive, acid-fast filaments were observed growing out from within these macrophage aggregates. The macrophage response seemed dependent upon the strain of Nocardia infecting them, since N. asteroides 10905 did not induce a similar response within the macrophage population.
Bacteriocin typing by leakage of ultraviolet light-absorbing material
Authors: Farkas-Himsley H , Pagel A
Published: January 2020

A rapid and reproducible method of bacteriocin typing is described based on leakage of ultraviolet light-absorbing material (UVAM), detectable in supernatants of bacteriocin-sensitive cultures, by means of a spectrophotometer. The prerequisites for reproducible results, with nonsignificant fluctuations in standard error of the mean, are: a set of standardized bacteriocins, produced under defined conditions and of determined strength. These must interact with the unknown bacterial culture in suspension and at a given ratio in order to achieve an optimal multiplicity of interaction. Pyocin and colicin typing by the "scrape and streak" technique of Gillies (J. Hyg. 62:1-10, 1963) was compared with the UVAM leakage method in 275 tests; the two tests were found to be in good agreement for the strains tested.
Polyol Metabolism by a Caries-Conducive
Streptococcus
: Purification and Properties of a Nicotinamide Adenine Dinucleotide-Dependent Mannitol-1-Phosphate Dehydrogenase
Authors: Brown Albert T. , Bowles Robert D.
Published: January 2020

The mannitol-1-phosphate dehydrogenase (M1PDH) (EC 1.1.1.17) from
Streptococcus mutans
strain FA-1 was purified to approximately a 425-fold increase in specific activity with a 29% recovery of total enzyme units, using a combination of (i) streptomycin sulfate and ammonium sulfate precipitation and (ii) diethyl-aminoethyl-cellulose (DE-52), agarose A 0.5M, and agarose-nicotinamide adenine dinucleotide (NAD) affinity column chromatography. Polyacrylamide gel electrophoresis of the purified enzyme preparation showed a single protein component that coincided with a band of M1PDH activity. The enzyme had a molecular weight of approximately 45,000 and was stable for long periods of time when stored at −80°C in the presence of β-mercaptoethanol. Its activity was not affected by mono- or divalent cations, and high concentrations of ethylenedia-minetetraacetic acid were not inhibitory. The M1PDH catalyzed both the NAD-dependent oxidation of mannitol-1-phosphate and the reduced NAD (NADH)-dependent reduction of fructose-6-phosphate. The forward reaction was highly specific for mannitol-1-phosphate and NAD, whereas the reverse reaction was highly specific for NADH and fructose-6-phosphate. The
K

m

values for mannitol-1-phosphate and NAD were 0.15 and 0.066 mM, respectively, and the
K

m

values for fructose-6-phosphate and NADH were 1.66 and 0.016 mM, respectively. The forward and reverse reactions catalyzed by the M1PDH from
S. mutans
appeared to be under cellular control. Both adenosine 5′-triphosphate and fructose-6-phosphate were negative effectors of the forward reaction, whereas adenosine 5′-diphosphate served as a negative effector of the reverse reaction catalyzed by the enzyme.
Interactions of radio-detoxified Escherichia coli endotoxin preparations with the complement system
Authors: Füst G , Bertók L , Juhász-Nagy S
Published: January 2020

Escherichia coli O89 lipopolysaccharide (LPS) was treated with different doses of gamma irradiation (5, 10, 15, and 20 Mrad). Various biological activities such as lethal effect, decrease in arterial blood pressure in dogs, and interaction with the complement system were determined for the parent and irradiated preparations. Irradiation of LPS significantly and in a dose-dependent manner decreased its lethal and blood pressure-depressing effects along with its ability to activate the complement system. In contrast, radio-detoxified LPS fixed more strongly the isolated human C1 than did the parent LPS. The possible connection between the toxicity of endotoxin and endotoxin-induced complement activation is discussed.
Polymyxins as inhibitors of polyclonal B-cell activators in murine lymphocyte cultures
Authors: Butler T , Smith E , Hammarström L , Möller G
Published: January 2020

The polymyxin antibiotics polymyxin B sulfate and colistin methane sulfonate were examined for their ability to inhibit responses to the polyclonal B-cell activators (PBA) bacterial lipopolysaccharide (LPS), dextran sulfate (DS), pneumococcal polysaccharide (SIII), and purified protein derivative of tuberculin (PPD) in spleen cell cultures. Polymyxin concentrations of 1 and 10 microng/ml significantly inhibited both the deoxyribonucleic acid synthetic and polyclonal antibody responses stimulated by LPS, DS, and SIII. At these concentrations of polymyxins, responses to PPD and to the T-cell mitogens concanavalin A and phytohemagglutinin were not affected. Inhibition was not caused by a generalized lymphocyte toxicity. After dialysis of LPS-polymyxin and DS-polymyxin mixtures, the PBA preparations showed decreased mitogenic activity. Thus, the polymyxins probably interacted directly with the LPS and DS molecules. The mitogenic response to DS was more significantly inhibited than the response to a nonsulfated dextran. The cationic property of the polymyxins probably allows attachment to negatively charged groups in the mitogenically relevant parts of some but not all PBA molecules,, this attachment resulting in the loss of PBA activity.
Homogeneity of protein serotype antigens in Neisseria meningitidis group A
Authors: Sippel J E , Quan A
Published: January 2020

Serotype antigens (STA), which have been shown to be constituents of outer-membrane protein, were extracted from group A meningococci with 0.2 M LiCl and pelleted by centrifugation at 150,000 X g. The STAs from 100 group A strains, which had been isolated from cases and carriers in various geographical locations, were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Every STA produced three major protein bands with molecular weights of approximately 35,000, 39,000, and 45,000, respectively. This SDS-PAGE pattern is clearly distinct from that produced by the serotype of group B and C meningococci most commonly isolated from cases (group B type 2 and group C type 2). The group A STAs were also indistinguishable by immunodiffusion. However, differences in bactericidal reactions were demonstrated, suggesting that there are other antigens that play a role in antibody response.
Streptococcus mutans
Dextransucrase: Functioning of Primer Dextran and Endogenous Dextranase in Water-Soluble and Water-Insoluble Glucan Synthesis
Authors: Germaine Greg R. , Harlander Susan K. , Leung Woon-Lam S. , Schachtele Charles F.
Published: January 2020

The extracellular enzyme activities of
Streptococcus mutans
6715 that synthesize glucans from sucrose were concentrated and partially purified by ammonium sulfate precipitation and gel permeation column chromatography. Polyacrylamide gel analysis demonstrated that all of the major proteins precipitated by ammonium sulfate were quantitatively recovered in the high-molecular-weight, enzyme-containing aggregates found in the void volume of the gel column. Anion-exchange column chromatography was used to fractionate the aggregates into preparations, α and β, which produced water-insoluble and water-soluble glucans, respectively. Polyacrylamide gel analysis showed that α and β contained unique proteins and dextransucrase (EC 2.4.1.5) activities. Studies on the time course of glucan synthesis by α demonstrated that this enzyme preparation contained dextranase activity, which partially degraded nascent alcohol-insoluble glucan into alcohol-soluble products that were subsequently reincorporated into insoluble product. The β enzyme preparation contained no detectable dextranase activity. Mixing experiments in the absence of primer dextran demonstrated that the dextranase activity present in α could modify glucan production by β. CsCl density gradient analysis of product glucans demonstrated that exogenous primer dextrans were used as acceptor molecules by both the α and β enzyme preparations, and that water-soluble glucans synthesized by β could be converted into water-insoluble glucans by α. It is proposed that the structural heterogeneity of the native glucans produced from sucrose by
S. mutans
is a result of the concerted action of glucan-forming dextransucrases and endohydrolytic dextranase activity.
Susceptibility of murine transfer factor to dimerized ribonuclease A
Authors: Rifkind D , Frey J A , Dinowitz M
Published: January 2020

Dialyzable transfer factor was prepared from the spleens of CF1 mice actively sensitized with killed Coccidioides immitis antigen. The transfer factor was administered to normal mice either intraperitoneally or into the hind footpads. The recipient mice were tested for reactivity to the coccidioides antigen and to Candida albicans antigen by means of the footpad swelling test. The transfer factor conferred antigen-specific reactivity upon normal recipient mice when given by the intraperitoneal and footpad routes. This capacity of the transfer factor was destroyed by in vitro pretreatment with dimerized ribonuclease A, an enzyme active against double-stranded, as well as single-stranded, ribonucleic acid. In contrast, monomeric ribonuclease A, which is active against only single-stranded ribonucleic acid under the conditions used here, was without effect upon the transfer factor. These data provide evidence that murine transfer factor contains ribonucleotides that are essential for immunological activity. In addition, the data are consistent with the hypothesis, advanced by others, that the ribonucleotides may be double-stranded or uniquely looped configurations.
Comparison of Coccidioides immitis arthrospore, mycelium, and spherule cell walls, and influence of growth medium on mycelial cell wall composition
Authors: Wheat R W , Tritschler C , Conant N F , Lowe E P
Published: January 2020

Comparative lipid content, cell wall yield, neutral monosaccharide, glucosamine, and protein (amino acid) contents of arthrospores, mycelia, and spherules of Coccidioides immitis Cash were studied. Cellular lipid contents were found in the decreasing order: spherules, arthrospores, mycelia. Lipid content of mycelia did not reach the level of arthrospores or spherules even when mycelia were grown on relatively rich media. Cell wall yields of spherules were lower than for mycelia when grown on comparable media. Cell walls of arthrospores, mycelia, spherules, and spherule culture filtrate all contained 3-O-methylmannose, mannose, and glucose, but in varying amounts. Cell wall yield and cell wall glucose content increased in mycelia grown in increasingly rich media, whereas mannose content either decreased or remained constant.
Classical and alternative complement pathway activation by pneumococci
Authors: Stephens C G , Williams R C , Reed W P
Published: January 2020

Sixty-two strains of Streptococcus pneumoniae were studied for their abilities to consume selected components of classical and alternative complement pathways in human sera. The classical pathway was blocked by chelating calcium with ethyleneglycol-bios (beta-aminoethyl ether)-N,N-tetraacetic acid and by removing C4. The alternative pathway was blocked by removing factor B. Each strain's activation of the two pathways was compared with its nonimmune reactivity with the Fc region of immunoglobulin G (IgG). Activation of the classical complement pathway appeared to be independent of such Fc reactivity. Highly Fc-reactive strains, however, were shown to activate the alternative pathway more effectively than did less Fc-reactive strains. Since pneumococcal activation of the alternative pathway requires non-immunospecific IgG, these findings suggest that nonimmune binding of IgG on the pneumococcal surface endows it with complement-activating properties.
Activation of botulinum toxins in the absence of nicking
Authors: Ohishi I , Sakaguchi G
Published: January 2020

The derivative toxins purified from cultures of proteolytic strains of Clostridium botulinum types A and F were found to have been only partially nicked but were fully activated. Trypsinization of C. botulinum type B derivative toxin at pH 6.0 resulted in simultaneous activation and nicking, whereas at pH 4.5, activation preceded nicking. The toxin was split by trypsin at pH 6.0 into two fragments with molecular weights of 112, ooo and 57,000. The toxin contained at least three trypsin-sensitive peptide bonds, one of which was more sensitive than the others at pH 6.0. These results indicate that activation of botulinum toxins by trypsin or endogenous protease (s) is not a direct result of nicking.
Comparative evaluation of human leukocyte and fibroblast interferon in the prevention of herpes simplex virus keratitis in a monkey model
Authors: Neumann-Haefelin D , Sundmacher R , Skoda R , Cantell K
Published: January 2020

The prophylactic potency of topically applied human leukocyte interferon and human fibroblast interferon was compared in a monkey model with herpes simplex virus type 1-induced keratitis. Both interferons effectively prevented keratitis at 1.9 X 10(6) U of activity per ml but were ineffective at 1.9 X 10(3) U/ml. Slight differences between human leukocyte interferon and human fibroblast interferon observed at the level of 1.9 X 10(5) U/ml were not statistically significant.
Regulation of invertase of Actinomyces viscosus
Authors: Kiel R A , Tanzer J M
Published: January 2020

The regulation of diethylaminoethyl-partially purified invertase (EC 3.2.1.26; beta-D-fructofuranoside fructohydrolase) from the 37,000 X g-soluble intracellular fluid of Actinomyces viscosus serotype 2 strain M-100 was studied. Glycolytic intermediates, mono-, di-, and triphosphate nucleotides, inorganic phosphate, and various divalent cations were tested for regulatory effects. Fructose-6-phosphate (F6P) and fructose-1,6-diphosphate (FDP) were found to act as noncompetitive inhibitors of invertase. The Ki values for F6P and FDP were found to be 3.4 and 5.1 mM, respectively. The Hill coefficient for sucrose was 1.03 and remained unchanged in the presence of varying amounts of F6P or FDP.
Microtiter solid-phase radioimmunoassay for detection of Escherichia coli heat-labile enterotoxin
Authors: Greenberg H B , Sack D A , Rodriguez W , Sack R B , Wyatt R G , Kalica A R , Horswood R L , Chanock R M , Kapikian A Z
Published: January 2020

The development of a microtiter solid-phase radioimmunoassay for the detection of Vibrio cholerae enterotoxin and heat-labile Escherichia coli enterotoxin is described. The test is based on the immunological similarity between V. cholerae toxin and E. coli heat-labile toxin. The assay is easy to perform, quantitative, and at least as sensitive and specific as the Y-1 adrenal cell system.
Expression of hepatitis B virus-specific markers in asymptomatic hepatitis B surface antigen carriers
Authors: Arnold G H , Shih J W , Kaplan P M , Purcell R H , Gerin J L , Bueschenfelde K H
Published: January 2020

A study was undertaken to assess the state of hepatitis B virus infection in a group of asymptomatic hepatitis B surface antigen (HBsAg) carriers. This study confirmed that the presence of hepatitis B e antigen (HBeAg) in serum was closely associated with serum HBsAg-specific deoxyribonucleic acid polymerase activity, hepatitis B core antigen (HBcAg) in serum and liver cell nuclei, and a histological picture of chronic hepatitis. No HBsAg-specific deoxyribonucleic acid polymerase activity or HBcAg was detected in highly concentrated anti-HBe-positive sera. In addition, liver biopsy specimens from carriers with anti-HBe were negative for HbcAg by immunofluorescence, and the liver histology was either normal or revealed only fatty changes. These data indicate that the anti-HBe-positive sera contained either no Dane particles or, if present, at least a 500-fold-lower concentration of Dane particles than that found in HBeAg-positive sera.
Rickettsial hemolysis: adsorption, desorption, readsorption, and hemagglutination
Authors: Winkler H H
Published: January 2020

The energy-dependent adsorption of radioiodinated rickettsiae to sheep erythrocytes was demonstrated. The iodination procedure, however, decreased the hemolytic activity of the rickettsiae. No desorption of rickettsiae from isolated rickettsia-erythrocyte complexes (prevented from lysing by NaF) could be measured. On the other hand, rickettsiae desorbed from this complex during or after lysis and readsorbed and lysed other erythrocytes. Thus, the usual hemolytic assay measures multiple rounds of adsorption and lysis. Although lysis of the rickettsia-erythrocyte complex was insensitive to anti-rickettsial rabbit serum, adsorption and readsorption were completely inhibited by such antiserum. Hemagglutination of erythrocytes by rickettsiae was observed (in the presence of NaF to prevent lysis) and was sensitive to the same inhibitors as adsorption.
In vivo studies on the cellular source of migration inhibitory factor in mice with delayed hypersensitivity
Authors: Salvin S B , Sonnenfeld G , Nishio J
Published: January 2020

Mice sensitized intravenously with 300 microgram of BCG cell walls in Drakeol-Tween 80 and challenged intravenously 3 weeks later with 50 mg of old tuberculin released migration inhibitory factor (MIF) into the circulating blood in quantities that could be detected in serum dilutions of 1:64 to 1:128. When thymus-derived lymphocytes (T-cells) were absent at the time of sensitization, as in neonatally thymectomized mice or in athymic nude mice, detectable amounts of MIF were not formed. Sensitized mice treated with either anti-theta serum or anti-bone marrow-derived lymphocyte (B-cell) serum before intravenous challenge with old tuberculin released reduced amounts of MIF into the circulation. Mice lethally irradiated, reconstituted with B-cells, sensitized with BCG cell walls, and then challenged intravenously 3 weeks later with old tuberculin did not release MIF into the circulation. When T-cells were injected at least 10 days before challenge, however, MIF appeared.
Factors influencing the appearance of antibody in tracheal washes and serum of young chickens after exposure to Newcastle disease virus
Authors: Ewert D L , Eidson C S , Dawe D L
Published: January 2020

The development of plaque-neutralizing antibody in tracheal washes and hemagglutination inhibition antibody in serum was followed after intratracheal and intranasal or intramuscular inoculation of 1-, 14-, or 28-day-old chicks with a lentogenic strain of Newcastle disease virus (NDV). Serum antibody could be detected between 7 and 10 days after intratracheal and intranasal vaccination in birds either with or without maternal antibody to NDV. However, among the 1-day-old group only birds without maternal antibody showed an antibody response after intramuscular inoculation. All birds possessing either actively or passively acquired serum antibody showed a sharp rise and subsequent decline of anti-NDV activity in tracheal washes between 4 and 10 days after intratracheal or intranasal vaccination. Using radiolabeled chicken immunoglobulin injected intravenously as a tracer, it was shown that this initial peak of anti-NDV activity in tracheal washes could be accounted for by enhanced transudation of serum antibody. The transudation of serum antibody coincided with the course of viral pathology observed in the tracheae of infected birds. Neutralizing antibody in tracheal washes beyond 10 to 14 days postvaccination was, most likely, porduced locally, in the respiratory tract.
Role of bacterial phospholipases in serum-mediated killing of Escherichia coli
Authors: Kreutzer D L , Vandermaten M , Buller C S , Robertson D C , Hirata A A
Published: January 2020

The importance of bacterial phospholipases during serum-mediated killing of Escherichia coli was examined by using wild-type DR+ DS+ and an isogenic phospholipase-deficient mutant DR- DS-. No difference in serum sensitivity was observed when the parental DR+ DS+ and mutant DR- DS- strains were exposed to various concentrations of normal guinea pig serum. Examination of the free fatty acid (FFA) and lipid composition during serum-mediated killing of the two E. coli strains indicated that FFA release occurred only in the parental DR+ DS+ strain. No FFA release or lipid degradation was detected in the mutant DR- DS- strain during serum killing. The addition of heat-inactivated E. coli antiserum (rabbit) to normal guinea pig serum caused FFA release in both E. coli strains. This FFA release was found to be independent of serum-mediated killing and due to a highly active and heat-resistant rabbit serum phospholipase that hydrolyzed the bacterial lipids after serum killing. The data presented indicate that serum-mediated killing of E. coli is independent of FFA release and that activation of bacterial phospholipases and the resulting release of FFA are only a result rather than a cause of serum-mediated cell death.
Arginine catabolism by Mycoplasma meleagridis and its role in pathogenesis
Authors: Ibrahim A A , Yamamoto R
Published: January 2020

A thin-layer chromatography technique was used to study the arginine metabolism of Mycoplasma meleagridis. The technique reflected the enzyme activity of the dihydrolase pathway through detection of readily visible end products on X-ray film. Strains of M. meleagridis differing in their pathogenicity for turkeys did not vary in arginine metabolism. In addition, no significant difference was observed in plasma arginine concentrations between M. meleagridis-infected and uninfected poults. It was concluded that the pathogenesis of M. meleagridis infection in turkeys was not based on its competition with the host for arginine.
Cell-mediated immunity to tumor antigen in Marek's disease: susceptibility of effector cells to antithymocyte serum and enhancement of cytotoxic activity by Vibrio cholerae neuraminidase
Authors: Sharma J M
Published: January 2020

Spleen cells from chickens inoculated 7 to 8 days previously with Marek's disease virus were cytotoxic for 51Cr-labeled cells of a Marek's disease lymphoblastoid cell line (MSB-1 line) in a 4-h in vitro cytotoxic assay. The cytotoxic activity of spleen cells was inhibited by pretreatment with antithymocyte serum and complement, but not with complement alone or in combination with anti-bursa cell serum or normal preimmune serum. The conclusion was that the effector cell in the above cytotoxic assay was a thymus-derived lymphocyte. Also, pretreatment of target cells with Vibrio cholerae neuraminidase enhanced in vitro cytotoxic activity of effector cells. Similar enzymatic treatment of effector cells had a negligible effect on cytotoxicity.
Quantitative immunoelectrophoretic assay for murine oncornavirus p30: noncovalent facilitation by sodium dodecyl sulfate
Authors: Robinson O R , Shibley G P , Sevoian M
Published: January 2020

Treatment of Rauscher murine leukemia virus lysates with the anionic detergent sodium dodecyl sulfate (SDS) at concentrations between 0.2 to 2.0% SDS per mg of viral protein greatly increased the anodal electrophoretic mobility of p30, the major internal polypeptide. SDS treatment did not reduce p30 antigenicity or cause nonspecific precipitation of normal serum proteins during subsequent immunoanalysis. The increased anodal electrophoretic mobility allowed assay of Rauscher murine leukemia virus p30 by Laurell rocket immunoelectrophoresis. An SDS-facilitated rocket immunoelectrophoresis assay is described that was highly reproducible (coefficient of variability, less than 3.0%) and capable of detecting 125 ng of viral protein. To our knowledge, this is the first report of a quantitative immunoelectrophoretic assay for an oncornavirus antigen. Since SDS binding is a general property of proteins, this method of noncovalently altering electrophoretic mobility appears to be applicable to other antigen-antibody systems.