bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased adhesion of Chinese hamster ovary cells to substratum by cholera enterotoxin
Authors: Nozawa R , Kon F , Yokota T , Ohashi M , Kuwahara S
Published: January 2020

A new, simple assay method using Chinese hamster ovary cells was devised for cholera enterotoxin. The effect of the cholera toxin was measured by an increase in cell adhesion to the substratum. The increase was dependent on the concentration of the toxin used and was effective for a longer period of time than that of adhesion increased by dibutyryl cyclic adenosine monophosphate and theophyline. There was a delay of about 60 min before the increase in adhesion, using the toxin, appeared, whereas the increase caused by dibutyryl cyclic adenosine monophosphate appeared almost at once.
Blastogenesis as an in vitro correlate of delayed hypersensitivity in guinea pigs infected with Listeria monocytogenes
Authors: Fulton A M , Dustoor M M , Kasinski J E , Blazkovec A A
Published: January 2020

Randomly bred guinea pigs of both sexes were injected intracardially with one-half a 50% lethal dose of Listeria monocytogenes. When these animals were skin tested with 30 mug of a water-soluble extract of sonically disrupted Listeria, animals had uniformly detectable levels of delayed-type hypersensitivity (DTH) 6 days after infection. Histological examination of skin test reaction sites, after fixation in Helly fixative and Giemsa staining, revealed a classical tuberculin-type infiltrate consisting primarily of mononuclear cells with few polymorphonuclear cells. Many of the small vessels showed perivascular cuffing. When purified peritoneal exudate lymphocytes from these animals were cultured in vitro in the presence of various concentrations of Listeria antigen, it was fount that the optimal antigenic dose for specific antigen-induced incorporation of [3H]thymidine varied for individual animals. In contrast to the early onset of uniformly detectable levels of in vivo DTH, in vitro lymphocyte blastogenesis was not uniformly demonstrable until 14 days postinfection and remained highly significant on days 21, 28, and 84 postinfection. At 7 days postinfection, lymphocytes from 7 of 17 animals were capable of undergoing sifnificant blastogenesis. The Listeria antigen preparation was not mitogenic for peritoneal exudate lymphocytes from normal animals. It was found that no direct correlation exists between the in vivo levels of DTH and in vitro blastogenesis. Cell donors showing significant in vitro blastogenesis nevertheless were also skin test positive for most animals tested. Humoral antibody was found to play no significant role in the immune response of guinea pigs to a primary infection with Listeria monocytogenes.
Characterization of monkey peripheral neutrophil granules during infection
Authors: Rausch P G , Canonico P G
Published: January 2020

Rhesus monkey (Macaca mulatta) neutrophils were shown to contain the azurophilic granule maker enzymes myeloperoxidase and beta-glucuronidase but were deficient in the specific granule markers alkaline phosphatase (AKP) and lysozyme. Isopycnic centrifugation of leukocyte homogenates on linear sucrose gradients resulted in cosedimentation of myeloperoxidase and beta-glucuronidase with an equilibrium density of 1.18. After an intravenous inoculation of monkeys with Salmonella typhimurium AKP activity became marked, whereas that of beta-glucuronidase decreased and myeloperoxidase remained unchanged. Lysozyme was undetected throughout the course of the experiment, but was present in oil-induced peritoneal macrophages and peripheral mononuclear cells. The induced AKP exhibited partial latency and had an equilibrium density of 1.15. It is unclear, however, whether the induced AKP is associated with specific granules or cytoplasmic membranes. Hence, while these data are consistent with the presence of azurophilic granules in polymorphonuclear neutrophils from infected monkeys, the presence of specific granules in polymorphonuclear neutrophils of both uninfected and infected monkeys remains moot.
Immunological properties of glycolipids from membranes of Acholeplasma laidlawii
Authors: Ryan M D , Noker P , Matz L L
Published: January 2020

Glycolipids, the predominant class of lipids in the membranes of Acholeplasma laidlawii, are the haptenic determinants that react with anti-A. Laidlawii serum to fix complement. The predominant complement-fixing activity of the membrane glycolipids was associated with the monoglucoysyl diglyceride, diglucosyl diglyceride, glycerlphosphoryl diglucosyl diglyceride (GPDD), and an unknown lipid B, which did not react with ninhydrin but release glucose and glycerol and traces of phosphorus upon hydrolysis. The glycolipids monoglucosyl diglyceride and diglucosyl diglyceride or GPDD and unknown lipid B were paired as a result of their cross-reactions with selective antisera prepared with the aid of reconstituted membrane complexes containing membrane lipids. Reconstituted membrane complexes assembled from [14C]monoglucosyl diglyceride and delipidated membrane proteins gave optimal complement fixation titers before saturation of the complexes with the ]14C]monoglucosyl diglyceride. The phosphoglycolipid of the membrane, GPDD, was anticomplementary as a pure lipid, a cholesterol liposome, and a reconstituted membrane complex. This anticomplementary activity, which was caused by 3 mug of pure GPDD, affected both human and guinea pig complement. Although human C1, C4, C3, and C5 were not inhibited by GPDD, C2 was inhibited 10-fold by reconstituted membrane complexes containing 150 mug of GPDD. A role for this phosphoglycolipid is discussed in the hypothetical mechanism of inhibition of C2 attachment to SAC1, 4 sites.
Mitogenicity of formalinized toxoids of staphylococcal enterotoxin B
Authors: Spero L , Leatherman D L , Adler W H
Published: January 2020

Staphylococcal enterotoxin B is a potent mitogen for mouse and human lymphocytes. Mitogenic activity was retained after detoxification of the enterotoxin by formaldehyde at pH 5.0, 7.5, OR 9.5. The most active toxoid (pH 7.5) was separated into a monomeric, a dimeric, and a polymeric fraction (1 x 10(5) to 3 x 10(5) molecular weight) by gel filtration, and although each fraction demonstrated mitogenic activity, the polymeric fraction was clearly the most efficacious. These data show that mitogenicity of staphylococcal enterotoxin B does not depend on toxicity. This suggests that the mitogenic and toxic activities are effected by different sites on the molecule.
Enterotoxigenic intestinal bacteria in tropical sprue. IV. Effect of linoleic acid on growth interrelationships of Lactobacillus acidophilus and Klebsiella pneumoniae
Authors: Mickelson M J , Klipstein F A
Published: January 2020

The factors responsible for colonization of the small intestine by enterotoxigenic coliform bacteria in Puerto Ricans with tropical sprue are unknown, but epidemiological observations have suggested that they may be related to an increased dietary intake of long-chain unsaturated fatty acids, particularly linoleic acid, which is known to exert an inhibitory effect on the growth of gram-positive organisms that normally comprise the flora of the small intestine. We have examined, by using a glucose-limited continuous-culture system, what effect this fatty acid exerts on the growth relationships of enteric gram-positive and coliform bacteria. In this system, colonization by an invading strain of Klebsiella pneumoniae was prevented by the presence of an established culture of Lactobacillus acidophilus, principally by virtue of a lowered pH of the medium that was incompatible with Klebsiella growth. However, when the population density of L. acidophilus was reduced by the presence of a sufficient concentration of linoleic acid, the invading K. pneumoniae successfully colonized the system and, once established, suppressed the growth of L. acidophilus. These observations indicate that, under the conditions of our chemostat, gram-positive enteric bacteria suppress coliform growth and that this effect is reversible by the presence of linoleic acid. It remains to be established, however, what pertinence these in vitro observations have to conditions within the intestinal tract of persons living in the tropics.
Effect of anti-herpesvirus drugs on human and bovine lymphoid function in vitro
Authors: Babiuk L A , Rouse B T
Published: January 2020

Lymphocyte proliferative responses to phytohemagglutinin, pokeweed mitogen, and herpesvirus (herpes simplex virus and infectious bovine rhinotracheitis virus) antigens were evaluated in the presence of various concentrations of the anti-herpesvirus drugs, methylmethoxydeoxyuridine (OCH3CH2UdR), cytosine arabinoside was inhibitory at 25 mug/ml and cytosine arabinoside was inhibitory OCH3CH2UdR per ml, lymphocyte proliferative responses were unaffected and were not abolished even by concentrations of 2,500 mug/ml. In contrast, adenine arabinocide was inhibitory at 25 mug/ml and cytosine was inhibitory at 5 mug/ml. Both direct and antibody-dependent lymphocyte cytotoxicity of herpesvirus-infected target cells were highly resistant to OCH3CH2UdR; concentrations of 1,000 mug/ml were only marginally inhibitory. The anti-herpesvirus activity of all three drugs is similar (0.5 to 8 mug/ml) (L.A. Babiuk, B. Meldrum, V.S. Gupta, and B. T. Rouse, submitted for publication). The drug OCH3CH2UdR should be given a careful, well-controlled evaluation of its effectiveness in the treatment of herpes simplex infections in animals, perhaps including humans, since our results using in vitro models show that cellular responses important for recovery from herpesvirus infections are unaffected by doses far in excess of those found be be antiviral.
Comparison of the rate of absorption and proteolysis of [14C]choleragen and [14C]bovine serum albumin in the rat jejunum
Authors: Strombeck D R , Harrold D
Published: January 2020

[14C]choleragen was used to study the rate of disappearance of choleragen enterotoxin from the jejunum of rats. [14C]bovine serum albumin (BSA) was studied in a similar manner. Almost one-third of the labeled toxin had disappeared from the intestine after 6 h. Its rate of disappearance was the same in germfree rats as in conventional rats. The rate of proteolysis of [14C]choleragen and [14C]BSA by intestinal mucodal lysosomal enzymes was also studied. Neither was significantly degraded by neutral proteases; however, heat-inactivated toxin was. They were all degraded by acid proteases; however, the rate of BSA proteolysis was only one-third of that of toxin. Soybean trypsin inhibitor had no effect on the in vivo disappearance of toxin nor on the acid proteases. It did inhibit the neutral protease digestion of heat-treated toxin. Aprotinin and protamine inhibited disappearance in loops of gut but had no effect to inhibit degradation rates. Gangliosides inhibited both rates of disappearance and proolysis of toxin. These agents had some different effects on disappearance rates and proteolysis of BSA. The data indicate that cholera enterotoxin is absorbed by intestinal mucosal cells and is degraded by acid proteases in the cells.
Increased cyclic adenosine 3',5'-monophosphate content in guinea pig ileum after exposure to Staphylococcus aureus delta-toxin
Authors: O'Brien A D , Kapral F A
Published: January 2020

To compare Staphylococcus aureus delta-toxin with cholera toxin, which is known to increase cellular cyclic adenosine 3',5'-monophosphate (cAMP), studies were undertaken to determine the effect of delta-toxin on the cAMP content of guinea pig ileum maintained in vitro. Concentrations of delta-toxin as low as 0.40 mug/ml increased cAMP levels in guinea pig ileum after 2 h of incubation. Histological damage was seen in ileum exposed for 2 h to delta-toxin concentrations of 100 mug/ml. As little as 3 mug of delta-toxin increased vascular permeability in guinea pig skin. Permeability changes became evident within 5 min and were maximal within 6 h, whereas those produced by cholera toxin required 24 h to become maximal. Benadryl did not interfere with the ability of these toxins to alter vascular permeability. Purified egg lecithin reduced the effectiveness of delta-toxin in the skin but did not inhibit cholera toxin. Delta-toxin in concentrations as low as 0.1 mug/ml caused dislodgement of HeLa cells in tissue cultures. Therefore, delta-toxin appears unique in being the only bacterial toxin, currently known to alter water absorption in the ileum, that is capable of both increasing cAMP levels and being cytotoxic. These findings suggest a possible role for delta-toxin in the pathogenesis of staphylococcal enteritis.
Response of Neisseria gonorrhoeae to Bdellovibrio species
Authors: Drutz D J
Published: January 2020

Bdellovibrio species are small, highly motile bacteria that are predators upon other bacteria in nature. Bdellovibrios attach to, penetrate, replicate within, and destroy prey that share the general characteristic of gram negativity. The lipopolysaccharide moiety of the cell membrane of target microorganisms appears to contain the principal receptor site for bdellovibrio attachment. Since gonococci also contain lipopolysaccharide that is similar in many respects to that contained within gram-negative rods, studies were conducted to determine the extent of gonococcal interaction with a variety of bdellovibrio species. Despite transient attachment, penetration of gonococci by bdellovibrios never occurred. Failure of bdellovibrio parasitization was unrelated to gonococcal species, colony type, piliation, penicillin susceptibility, or virulence as influenced by passage in embryonated eggs. In experiments involving mixtures of gonococci and more typical gram-negative bacillary prey, the latter were always attacked by bdellovibrios, whereas the former were ignored. Despite evidence for similarities between gonococcal and gram-negative bacillary lipopolysaccharides, resemblances do not extend to the point where gonococci are susceptible to bdellovibrio parasitization.
Ganglioside and rabbit erythrocyte membrane receptor for staphylococcal alpha-toxin
Authors: Kato I , Naiki M
Published: January 2020

The hemolytic activity of staphylococcal alpha-toxin is inhibited by an N-acetylglucosamine-containing ganglioside (GlcNAc-ganglioside) but not by any of the related glycolipids. The GlcNAc-ganglioside also is precipitated with the toxin by a gel-diffusion technique. It is postulated that GlcNAc-ganglioside may be the membrane receptor for the alpha-toxin.
Effect of viral and bacterial pneumonias on cell-mediated immunity in humans
Authors: Kauffman C A , Linnemann C C , Schiff G M , Phair J P
Published: January 2020

Cell-mediated immunity (CMI) was assessed during infection and after convalescence in 12 patients with influenza pneumonia and 10 patients with bacterial pneumonia. The patients with influenza pneumonia had a marked impairment of skin test reactivity, and their lymphocytes showed a diminished response to phytohemagglutinin and streptokinase-streptodornase stimulation in vitro. Suppression of CMI was related to the severity of the pneumonia. Patients with bacterial pneumonia showed as great a suppression of the response to phytohemagglutinin and streptokinase-streptodornase as the patients with viral pneumonia. All parameters of CMI returned to normal in both groups after convalescence. The depression of CMI could not be related to a decrease in the number of thymus-derived lymphocytes or to serum-suppressive factors in these patients.
Hydrolysis of urea in the gastrointestinal tract of "monoxenic" rats: effect of immunization with strains of ureolytic bacteria
Authors: Moreau M C , Ducluzeau R , Raibaud P
Published: January 2020

Axenic rats, in whose feces urea is ordinarily excreted, were inoculated with ureolytic strains of Lactobacillus or Actinobacillus originally derived from the microflora of "holoxenic" rats. In these "monoxenic" animals, harboring one or another of the bacterial strains, fecal urea was hydrolyzed, with a more rapid onset of ureolysis in the case of Actinobacillus as compared with Lactobacillus. In vitro, a parallel difference between the two strains with regard to the onset of ureolysis was observed, hydrolysis beginning at the onset of growth in the case of Actinobacillus and only at the end of the exponential growth phase in the case of Lactobacillus. Extracellular urease activity was demonstrated in cultures of Lactobacillus, whereas none was found extracellularly with Actinobacillus. The pH optimum for the Lactobacillus urease in vitro was found to be 3.0, whereas the corresponding value for Actinobacillus was 6.0. In the two types of monoxenic rats, urea was consistently present in the small intestine and virtually absent from cecum and colon. Hydrolysis of urea in stomach was almost complete in rats bearing Lactobacillus but much less so in animals monoxenic with Actinobacillus, despite essentially equal numbers of organisms in that location. When rats carrying a monoflora of ureolytic Lactobacillus were immunized with either whole cells or soluble extract of the same organism, urea appeared in cecum and feces, indicating suppression of ureolytic activity. Immunization with an extract of nonureolytic Lactobacillus failed to produce such a result. Similar immunization techniques applied to animals monoassociated with ureolytic Actinobacillus did not alter ureolysis, and no appreciable quantity of urea appeared in feces. These studies demonstrate that it is indeed possible to inhibit the ureolytic activity of some bacteria in vivo by immunological means, but that the urease system of other organisms may not be as amenable to such manipulation.
Effect of biotin deficiency on some properties of Staphylococcus aureus isolates from humans and other animals
Authors: Adekeye J D , Coles E H
Published: January 2020

Staphylococcus aureus isolates from humans and other animals were grown in biotin assay medium containing 12 mug of biotin per liter and compared to isolates from the same sources grown concurrently in medium containing adequate biotin. The two cultures were tested for production of coagulase, phosphatase, and fibrinolysin enzymes and for responses to various antimicrobial agents and bacteriophages. Organisms grown in biotin-deficient medium produced less phosphatase; coagulase and fibrinolytic activity was reduced, and they were more susceptible to antimicrobial agents than were normal organisms, but phage susceptibility was not greatly affected.
Inhibitory action of D-galactose on phagocyte metabolism and function
Authors: Litchfield W J , Wells W W
Published: January 2020

To account for enhanced susceptibility to infection among galactosemics, the acute effects of D-galactose on metabolic and functional activities of phagocytic cells in vitro were investigated. Human and guinea pig polymorphonuclear leukocytes (PMN) when incubated in medium containing 30 mM galactose displayed substantially less killing of Escherichia coli than when incubated in medium with 5 mM glucose. Impaired bactericidal activity was dependent upon galactose concentration but could be partially averted by supplementing the galactose-containing medium with 15 mM glucose. Phagocytic activities of guinea pig PMN and peritoneal macrophages were assayed by following ingestion of 32P-labeled E. coli and were also depressed by elevated galactose. Galactose was readily epimerized to glucose by resting PMN, and this conversion was stimulated by phagocytosis. Incubation of macrophages in the presence of galactose resulted in depletion of intracellular levels of adenosine 5' -triphosphate as well as other metabolities.
Relationship of cellular potential hemolysin in group A streptococci to extracellular streptolysin S
Authors: Calandra G B , Whitt R S , Cole R M
Published: January 2020

The relationship of streptolysin S (SLS) and a cellular potential hemolysin (PH) activatable by sonication was examined in strain C203S (a known high producer of SLS), its SLS(-) mutant (C203U), and in 20 other group A streptococci of various M and T serotypes. All strains shown to form SLS (ribobycleic acid (RNA)-core SLS) contained PH. The two strains lacking PH were the only ones that did not produce SLS In strain C203S, formation of SLS bycells incubated with RNA-core for 60 min at 47 C in a nongrowth basal medium (Bernheimer's basal medium) was followed by a marked decrease (99.6% loss) of PH titer. Without stimulation of SLS production by addition of RNA-sore, the same incubation resulted in a progressive but less marked fall (38.8%loss in 60 min) of PH titer: these cells produced disproportionately low titers of SLS on subsequent addition of RNA-core. This effect of prior incubation in Bernheimer's basal medium on SLS titer was partially nullified by use of fresh medium after 30 min, but not after 60 min, and did not occur during 60 min of incubation at OC. These results provide additional evidence for a precursor-product relationship between PH and SLS. They also suggest that a medium factor (or factors) is utilized or destroyed at 37 C and that this factor is essential to both the stability of PH and its efficient conversion to SLS.
Erythrocyte antinuclear antibodies in sera of chickens hyperimmunized with group A streptococcal vaccine
Authors: Luster M I , Leslie G A
Published: January 2020

Chickens hyperimmunized with group A streptococcal vaccine often synthesize high levels of antibody to group A streptococcal carbohydrate (SACHO). Indirect immunofluorescent analysis revealed that sera of these chickens also contain antinuclear antibodies capable of reacting with chicken erythrocyte nuclei (EANA) at titers up to 2,560. Removal of the anti-SACHO antibodies from hyperimmune serum did not significantly reduce EANA titers, indicating that anti-SACHO antibodies are not responsible for the EANA reactions. The time course for antibody development as well as the immunoglobulin class distribution of EANA and anti-SACHO antibodies were very similar. Localization of EANA activity to Fab fragments indicated that the immunfluorescent reaction represents an antigen-antibody reaction. Findings point out an important consequence of using chicken immunoglobulin Y in immunofluorescent assays. The purified Fc fragment of immunoglobulin Y is cytophilic for the substrate when tested at high concentrations, which suggests that some cytophilic immunoglobulins are involved in the reaction.
Germination of Candida albicans induced by proline
Authors: Dabrowa N , Taxer S S , Howard D H
Published: January 2020

Blastospores of Candida albicans germinated in proline-biotin-buffer medium incubated at 37 C. Certain other amino acids in the glatamate, asparate, and pyruvate families also fostered germinaton but generally to a lesser extent than did proline. L-Cysteine, D-proline, and certain structural analogues of L-proline inhibited proline-stimualted germination. The concentration of phosphate and glucose was crucial to amino acid-stimulated germination of C. albicans. Clinical isolates and stock cultures varied in their response to the germ tube-inducing activity of proline or other amino acids. The proline-buffer medium cannot be used in a diagnostic test for production of germ tubes by isolates of yeasts.
Antigenicity of converting phages obtained from Clostridium botulinum types C and D
Authors: Oguma K , Iida H , Shiozaki M , Inoue K
Published: January 2020

Phage conversion of toxigenicity in Clostridium botulinum types C and D was accomplished by using nontoxigenic strains and phages purified from plaques. Although the morphology of the converting phages seemed to be the same, they were divided into three groups on the basis of their conversion spectrum. The first group consists of phages obtained from toxogenic strains C-Stockholm and C-468. The second group consists of phages from strains D-1873 and C-203. The third group consists of phages from strains D-South African and D-4947. These converting phages were also classified into the same three groups by a neutralization test with specific antiphage sera. Cross-neutralization, however, was observed between phages belonging to group 1 and group 2,by both the neutralization test of converting ability and by a plaque experiment in which the surviving rates of phages were calculated after treatment with each antiphage serum. The antigenic differences among these converting phages should probably comprise one of the reasons for the existence of the specific infection spectrum in C. botulinum types C and D.
Purification of hepatitis A antigen from feces and detection of antigen and antibody by immune adherence hemagglutination
Authors: Moritsugu Y , Dienstag J L , Valdesuso J , Wong D C , Wagner J , Routenberg J A , Purcell R H
Published: January 2020

Hepatitis A antigen (HA Ag) was purified from feces collected during acute illness from patients with naturally occurring viral hepatitis, type A. Positive fecal specimens were identified by immune electron microscopy, but for detection of HA Agduring purification immune adherence hemagglutination (IAHA) and microtiter solid-phase radioimmunoassay were used. Isopycnic banding in cesium chloride, rate-zonal separation in sucrose, and preparative zonal electrophoresis were used in various combinations for successive purification, and the purified antigen was successfully used in a test for antibody by IAHA. Seronconversions to HA Ag were demonstrated by IAHA in 20 instances of hepatitis A virus infection, but in none of six cases of type B hepatitis or three cases of post-transfusion hepatitis unrelated to heaptitis A or B viruses, nor in two individuals without hepatitis. In addition, the temporal pattern of antibody development during type A hepatitis was studied in serial sera from an experimentally infected chimpanzee. Antibody titers by IAHA correlated well with antibody ratings determined by immune electron microscopy.
Lymphocytic choriomeningitis virus infection in fetal, newborn, and young adult Syrian hamsters (Mesocricetus auratus)
Authors: Parker J C , Igel H J , Reynolds R K , Lewis A M , Rowe W P
Published: January 2020

The pathogenesis of lymphocytic choriomeningitis virus infection in fetal, newborn, and young adult hamsters was studied. Infected newborn hamsters initially developed a persistent viremia and viruria with titers often in excess of 10(4.0) mean infectious doses/0.03 ml of blood or urine. After week 12 two different patterns of infection became evident. Approximately one-half of the hamsters eventually cleared the infection, whereas the others developed a chronic progressive and ultimalely fatal disease characterized by continuous high-titered viremia and viruria and high titers of virus in their tissues. Complement-fixing antibody and, to a lesser degree, virus-neutralizing antibody coexisted with the viremia. Hamsters with persistently high levels of viremia and viruria developed chronic glomerulonephritis and widespread vasculitis, whereas hamsters that cleared their infections did not develop these lesions. Litters of hamsters born to viremic mothers were invariably infected. Litter sizes were small and breeding effectiveness was reduce; however, vertical, congenital infection was successfully passed through three generations. The course of infection in the congenitally infected hamsters was similar to that in newborn infected hamsters, with all animals producing complement-fixing antibody, some animals being capable of clearing the viremia and remaining healthy, and other animals having persistent viremia and fatal disease. Inoculated young adult hamsters did not become diseased, developed viremia and viruria which persisted up to 3 and 6 months, respectively, and developed complement-fixing antibody by 10 days after infection. The prolonged urinary excretion of large amounts of lymphocytic choriomeningitis virus by asymptomatic, chronically infected hamsters is an important public health consideration when dealing with potential human infection.
Purification of staphylococcal alpha-toxin by adsorption chromatography on glass
Authors: Cassidy P , Harshman S
Published: January 2020

Staphylococcal alpha-toxin was purified from Staphylococcus aureus growth medium using adsorption chromatography on controlled pore glass beads. Elution of alpha-toxin from the unmodified glass surface of the beads with various anions generally followed the chaotropic series. Alpha-toxin, purified by glass bead chromatography, is composed of a single electrophoretic form, containing less than 2% of other forms.
Ontogeny of human cell-mediated immunity: age-related variation of in vitro infantile lymphocyte transformation
Authors: Alford R H , Cartwright B B , Sell S H
Published: January 2020

Thymidine uptake of unstimulated and of phytohemagglutinin (PHA)-, streptokinase-streptodornase (SK-SD)- and candida extract (candida)-stimulated lymphocytes of normal infants less than 20 months old was evaluated. Thymidine uptake of unstimulated and PHA-stimulated young infants' lymphocytes was avid, resembling that of newborn cells. Over periods of weeks to months, infantile lymphocytes demonstrated transition in unstimulated and PHA-induced thymidine uptake from response patterns like cord blood cells to ones more characteristic of adult lymphocytes. Candida- and SK-SD-induced thymidine uptake of lymphocytes from very young infants was likewise apparently quite avid, resembling cord blood cells. However, factoring out high unstimulated thymidine uptake by conversion of data to stimulation indexes clarified differing paces of acquisition of transformation responsiveness of the two naturally acquired infectious antigens. Specific lymphocytes reactivity to SK-SD was quite low in all groups of infants compared with candida-induced transformation, which in some infants acquired adult proportions.
Dextran-mediated interbacterial aggregation between dextran-synthesizing streptococci and Actinomyces viscosus
Authors: Bourgeau G , McBride B C
Published: January 2020

Streptococcus sanguis and Streptococcus mutans bind to the surface of Actinomyces viscosus, producing large microbial aggregates. Aggregates form rapidly and are not easily dissociated by vigorous mixing. The binding is mediated by dextran. Glucose-grown streptococci will not aggregate unless they are first mixed with high-molecular-weight dextran. Aggregation is induced with dextrans isolated from Leuconostoc, S. sanguis, or S. mutans. Sucrose-grown streptococci will adhere to A. viscosus without the addition of an exogenous source of dextran. A. viscosus will bind dextran and then bind glucose-grown streptococci. Aggregation occurs over a wide pH range and is dependent on cations. The aggregating activity of A. viscosus is both protease and heat sensitive. The aggregating activity of S. sanguis is heat stable but sensitive to dextranase.
Establishment of cutaneous Leishmania enriettii infection in hamsters
Authors: Belehu A , Turk J L
Published: January 2020

A model of a self-healing type of cutaneous leishmaniasis was established in hamsters using the guinea pig parasite Leishmania enriettii. L. enriettii was passaged several times in hamsters without losing its infectivity for guinea pigs or for hamsters. The course of the infection in hamsters resembled that of guinea pigs, with the exception that the lesion at the site of parasite inoculation did not ulcerate and no metastatic lesions developed spontaneously. Moreover, unlike guinea pigs, infected or recovered hamsters were skin test unresponsive to various preparations of L. enriettii antigens. However, histological examination of draining lymph nodes showed features of a cell-mediated immune response, and in vitro inhibition of macrophage migration was demonstrable using peritoneal exudate cells from recovered animals and specific leishmanial antigen. Antibody was demonstrable by indirect immunofluorescence starting 1 week after infection. Recovered animals were immune to reinfection; however, the passive transfer of peritoneal exudate cells or serum from recovered animals did not confer protection against L. enriettii infection in normal animals.
Solid-phase radioimmunoassay for antibodies to flavivirus structural and nonstructural proteins
Authors: Trent D W , Harvey C L , Qureshi A , LeStourgeon D
Published: January 2020

A micro-solid-phase radioimmunoassay (SPRIA) is described for quantitation of antibodies to purified flaviviruses as well as to the purified envelope glycoprotein and 80,000-molecular-weight viral nonstructural protein. Sera from mice experimentally infected with Saint Louis encephalitis (SLE) virus or from humans after a primary SLE virus infection reacted more specifically with the major viral envelope protein in the SPRIA test than with antigens conventionally used in the complement fixation (CF) and hemagglutination inhibition tests. A high degree of correlation (P is less than 0.05) was observed between SPRIA anti-immunoglobulin G binding values with the 80,000-molecular-weight nonstructural protein of SLE virus and antibody titers obtained by plaque reduction neutralization and CF with the nonstructural protein. In five of seven human sera in which CF antibody titers to the nonstructural protein were 4 or less, SPRIA testing revealed significant titers of IgG immunoglobulin reactive with this viral protein. The SPRIA test for antibodies reactive with group B togavirus nonstructural protein is as specific and sensitive as the plaque reduction neutralization test for titrating viral antibody in human and animal sera. Antibodies reactive with viral envelope proteins are broadly cross-reactive by the Spria technique, demonstrating both group- and complex-reactive antigenic determinants. The SPRIA test, using wells precoated with antigen, can be completed in 1 day, providing a rapid, highly sensitive test which can be adapted to use in testing a large number of sera.
Immunity to syphilis: passive transfer in rabbits using serial doses of immune serum
Authors: Weiser R S , Erickson D , Perine P L , Pearsall N N
Published: January 2020

The influence of prolonged treatment with serial doses of immune serum initiated either 2 or 48 h before intracutaneous challenge with Treponema pallidum on the subsequent appearance and development of lesions was studied. Whereas treatment with large doses of immune serum initiated 48 h before treponemal challenge caused a delay of 9 to 20 days in the onset of lesions and marked suppression of lesion development. Treatment initiated 2 h before treponemal challenge failed to delay the onset of lesions and only moderately suppressed their development. Lesion development was suppressed as long as treatment was continued. The significance of the results is discussed.
Autoimmunity induced in rabbits by rinderpest virus
Authors: Fukuda A , Yamanouchi K
Published: January 2020

During rinderpest virus infection in rabbits, 19S cold hemagglutinating antibody against rabbit erythrocytes and 7S anti-nuclear antibody, which reacts with the nuclei and/or the nuclear membranes in immunofluorescent staining, were demonstrated. Virus infection that affected the thymus-dependent immune functions was speculated to act as a trigger for the production of these two autoantibodies.
Morphological and steroidogenic changes in cultured adrenal tumor cells induced by a subunit of cholera enterotoxin
Authors: Donta S T , Kreiter S R , Wendelschafer-Crabb G
Published: January 2020

A purified subunit of the cholera enterotoxin molecule was found to have morphological and steroidogenic inducing effects similar to those induced by the native enterotoxin on monolayer tissue cultures of Y1 adrenal tumor cells, although 1,000 times more subunit than toxin (weight basis) was required for maximal effects. In contrast to the whole toxin, the effects of the active subunit could not be prevented by prior incubation with either Gm1 ganglioside or with antibodies directed against choleragenoid (the binding subunit). These results suggest that different receptor sites may exist on cells for the binding and for the active subunits of cholera enterotoxin and/or that the active toxin fragment may exert its effects after gaining access to the intracellular compartment.
Serological studies with reovirus-like enteritis agent
Authors: Blacklow N R , Echeverria P , Smith D H
Published: January 2020

The age distribution of antibody to the human reovirus-like enteritis agent, and to the antigenically related Nebraska calf diarrhea virus (NCDV) was studied in serum specimens obtained from 592 children hospitalized in Boston. Sera were examined for complement-fixation (CF) antibody to the human agent, and for CF and indirect immunofluorescence-staining antibodies to NCDV. The curve of antibody frequency was similar in each of the three assays, showing a steep rise in the 6- to 18-month-old age groups; these results indicate the early acquisition of antibody to the reovirus-like enteritis agent. The majority of children and young adults possessed CF- and immunofluorescence-stainable antibodies. There was a significant association between antibody prevalence data obtained with the human CF antigen and with the two NCDV antigens; this association was closest between the human and NCDV CF antigens.
Endotoxic activity of rough organisms of Brucella species
Authors: Jones L M , Diaz R , Berman D T
Published: January 2020

A rough-specific antigen extracted from the rough species Brucella ovis and lipopolysaccharide extracted from smooth Brucella abortus demonstrated equivalent levels of activity in tests for mouse lethality and limulus lysate clotting activity. Acetone-extracted whole cells of B. ovis and of B. canis and of a rough mutant of B; abortus had the same toxicity for mice, but it was not possible to extract endotoxin from B. canis by the methods used.
Further characterization of a lipopolysaccharide from Coxiella burneti
Authors: Chan M L , McChesney J , Paretsky D
Published: January 2020

The lipopolysaccharide previously isolated from the rickettsial agent of Q fever, Coxiella burneti, phase I, has been further characterized. The sugar residues ribose, mannose, gluclose, D-glycero-D-mannoheptose, and L-glycerto-D-mannoheptose are present. Two sugars remain unidentified, one of which is a minor and the other a major constituent. Isomyristic, palmitic, and beta-hydroxymyristic acids are the major fatty acid residues of the 15 identified. The nature and content of other lipopolysaccharide constituents are presented.
Antigenic relationships on the diphtheria toxin molecule: antitoxin versus antitoxoid
Authors: Rittenberg M B , Pinney C T , Iglewski B H
Published: January 2020

We used the mouse to produce antisera to native diphtheria toxin and diphtheria toxoid. With these antisera it was possible to distinguish between toxin and toxoid. By gel diffusion analysis, antitoxin detected antigenic determinants on toxin which were not available on toxoid, indicating that some determinants had been lost or altered by formalin treatment. Antitoxoid, on the other hand, showed reactions of identity between toxin and toxoid in gel diffusion. The toxin neutralization titers measured in tissue culture were the same for both antisera. Only antitoxin neutralized the adenosine 5'-diphosphate ribosyl-transferase activity of fragment A, but suprisingly both antisera had significant anti-fragment A titers when tested by passive hemagglutination. It is suggested that some of the anti-fragment A activity in antitoxin affects the enzyme active site, whereas that in antitoxoid does not, implying the existence of a least two independent antigenic regions on fragment A.
Influence of feed on onset of disease in subacute myelo-optico-neuropathy virus-infected mice
Authors: Inoue Y K , Nishibe Y
Published: January 2020

An enriched feed, specially prepared to affect reproduction of mice, significantly suppressed the onset of disease in subacute myelo-optico-neuropathy virus-infected mice, whereas low-calorie feed enhanced the incidence of disease.
Cell-mediated immune response to products of Actinomyces viscosus cultures
Authors: Turner D W , Roberson B S , Longton R W
Published: January 2020

Hartley strain guinea pigs were sensitized with 0.5 ml of concentrated cell-free Actinomyces viscosus culture supernatant fluids mixed with Freund complete adjuvant. Fourteen to 16 days later the animals were challenged by intradermal injection with 0.1 ml of the culture supernatant, and the reactions were observed at 4, 8, 16, 24, and 48 h. Peritoneal exudate cells from sensitized animals were used for determination of migration inhibition factor, and guinea pig peripheral blood served as a source of cells for determining the induction of mitogenesis by antigenic material. Skin responses were consistently positive to challenge with the test material, whereas reactions to noninoculated culture medium were negative. Sensitized cells, challenged with antigen, resulted in 60% or greater inhibition of migration of indicator cells in migration inhibition factor experiments. Tests for mitogenesis showed a greater than fourfold increase in isotope uptake when sensitized cells were challenged with test material. The data are consistent with the suggestion that A. viscosus culture supernatants contain substances that induce cell-mediated immune responses in guinea pigs.
Rat polyvinyl sponge model for the study of infections: host factors and microbial proliferation
Authors: Isenberg H D , Wiener S L , Isenberg G A , Sampson-Scherer J , Urivetzky M , Berkman J I
Published: January 2020

Female rats were treated with several administration regimens of methylprednisolone, cobra venom anti-complementary factor, and cyclophosphamide in conjunction with polyvinyl sponge implantations. The effect of these drugs on host factors active against bacteria was evaluated with Staphylococcus aureus ATCC 25933, Escherichia coli K-12, and Pseudomonas aeruginosa CDC 7725. One of two implants in each animal was infected with 10(8) of one of the three bacteria, and bacterial and granulocyte content was determined in the infected and control sponges after 48 h. The single large dose of methylprednisolone decreased staphylococcal and E. coli clearance while promoting dissemination of P. aeruginosa. A low chronic dose of the steroid inhibited E. coli chemotaxis only. A higher dose of the steroid administered chronically interfered markedly with S. aureus and E. coli curtailment by the host while leading to enhanced dissemination of P. aeruginosa, accompanied by a precipitous decline in granulocytes. Results with cobra factor resembled the higher chronic dose of steroid enhancing, especially the dissemination of the pseudomonad and its anti-granulocytic propensity. Cyclophosphamide depression of granulocytes revealed the rat's ability to curtail the proliferation of particular S. aureus and E.coli strains even in the absence of leukocytes. This treatment resulted in the rapid spread of P. aeruginosa, leading to the death of some experimental animals. These experiments underline the versatility of this animal model in the study of host and microbial factors influential in infectious disease.
Phage conversion to hemagglutinin production in Clostridium botulinum types C and D
Authors: Oguma K , Iida H , Shiozaki M
Published: January 2020

Five toxigenic strains of Clostridium botulinum types C and D were incubated at 37 degrees C for 7 days in 15 ml of the following media: LYG medium, cooked-meat medium, egg meat medium, and N-Z-amine medium. The supernatants of these cultures were tested for hemagglutinin production with 1% erythrocytes obtained from mice, guinea pigs, chickens, sheep, monkeys, and humans. Four toxigenic strains produced hemagglutinin. The highest hemagglutinin titer was obtained with a combination of human erythrocytes and cultures incubated in LYG medium. When the same experiment was carried out with many nontoxigenic strains, hemagglutination was observed in only one strain, C-N71. Strains producing hemagglutinin also produced phages. The phages obtained from toxin- and hemagglutinin-producing strains converted nontoxigenic indicator strains to produce both toxin and hemagglutinin. The phage obtained from a toxin-positive hemagglutinin-negative strain could only induce cultures to produce toxin, and the phage from a toxin-negative hemagglutinin-positive strain could only induce production of hemagglutinin. These studies suggest that the production of hemagglutinin by C. botulinum types C and D is governed by bacteriophages and that hemagglutinin production can be transmitted separately or concomitantly with toxin production.
Assay of Escherichia coli enterotoxins by in vivo perfusion in the rat jejunum
Authors: Klipstein F A , Lee C S , Engert R F
Published: January 2020

Assay of Escherichia coli enterotoxins by in vitro perfusion in rats was evaluated by examining the effects of variously prepared fractions of heat-labile (LT) and heat-stable (ST) toxins on water transport in this system. The assay was found to respond equally well, in a dose-related manner, to both LT and ST; it was sufficiently sensitive to detect the toxigenic effect of concentrations as small as 1 ng/ml. With the assay, it was found that LT is produced in cultures grown under aerobic, but not anaerobic, conditions; in contrast, ST is elaborated in stationary aerobic and anaerobic broth cultures but not in those grown under agitated aerobic conditions. Both toxins can be precipitated by either ammonium sulfate or acetone. The two toxin forms were completely separated from each other by sequential ultrafiltration. LT alone (thermolabile after exposure to 100 degrees C for 30 min) was retained by a PM-30 membrane, and ST alone was present in UM-2 retentates; ST was retained more effectively by a UM-05 membrane, with a 1,000-fold increase in activity over that of the UM-2 retentate. Washed ultrafiltration retentates containing either LT or ST derived from the proper culture conditions all induced water secretion at concentrations of 100 ng or less per ml. These results indicate that in vivo perfusion in rats is a sensitive, duplicable assay for both the LT and ST forms of E. coli enterotoxin.
Tumor inhibition in mice by lipopolysaccharide-induced peritoneal cells and an induced soluble factor
Authors: Berendt M J , Saluk P H
Published: January 2020

Lipopolysaccharide (LPS) was shown to prevent tumor growth in BALB/c mice when administered either prior to or after the inoculation of lethal doses of tumor cells. An attempt to elucidate the mechanism of this phenomenon utilizing in vivo protocols was made by the adoptive transfer of tumor protection with peritoneal cells as well as with cell-free peritoneal fluids obtained from non-tumor-bearing, LPS-stimulated syngeneic mice. The in vivo-activated peritoneal cells from LPS-treated mice were capable of adoptively transferring tumor protection at peritoneal cell to tumor cell ratios ranging from 1,000:1 to 100:1. Experiments were also performed that indicate that: (i) LPS exerts no direct toxic or inhibitory effect on the tumor cells, and (ii) that residual LPS present in cell and fluid preparations was not responsible for such protection.
Viral infections in renal transplant recipients
Authors: Armstrong J A , Evans A S , Rao N , Ho M
Published: January 2020

Cytomegalovirus (CMV) infections are common in renal transplant recipients. We studied 23 recipients prospectively to determine whether infections by other herpes-group and non-herpes-group viruses were also present. Sera, obtained at the time of surgery and periodically thereafter, were tested for antibody to CMV, herpes simplex virus (HSV), Epstein-Barr virus (EBV), parainfluenza viruses types 1, 2, and 3, and the viruses of measles and rubella. We found no evidence of an unusual incidence of primary or secondary infection by the non-herpesviruses tested. Rises to CMV, HSV, and EBV antibody titers occurred in 43, 38, and 32% of patients, respectively. All serological rises to herpes-group viruses occurred in patients seropositive at the time of transplantation, with the exception of three patients who experienced primary CMV infections. We conclude that reactivation of all herpes-group viruses tested may occur in transplant recipients. Morbidity was associated only with primarly CMV infection.