bluuebunny/crossref_metadata_2025 huggingface dataset:
Epicillin: Experimental Chemotherapy, Pharmacodynamics, and Susceptibility Testing
Authors: Gadebusch H. , Miraglia G. , Pansy F. , Renz K.
Published: January 2020

Epicillin exhibits excellent therapeutic activity when administered orally or subcutaneously to mice infected with a variety of pathogenic bacteria. These included single strains of
Streptococcus pyogenes
, non-penicillinase-producing
Staphylococcus aureus, Proteus mirabilis, Escherichia coli, Salmonella schottmuelleri
, and
Pseudomonas aeruginosa
, strains of microorganisms selected as typical of those that frequently cause serious clinical infections. In oral chemotherapeutic comparisons of tetracycline, chloramphenicol, cephalexin, ampicillin, and epicillin, the latter, with but minor exceptions, exhibited activity comparable to that of most of the other antibiotics studied. Oral administration to mice of a loading dose of epicillin or ampicillin resulted in similar peak blood levels and decay curves. The concentration in urine and the excretion rates were significantly higher in epicillin than in ampicillin-treated animals. Presumptive evidence suggests that the inter-pretation of zone diameters obtained with 10-μg epicillin discs should be the same as is used for 10-μg ampicillin discs.
Light and Scanning-Beam Electron Microscopy of Wall-Defective
Staphylococcus aureus
Induced by Lysostaphin
Authors: Watanakunakorn Chatrchai , Fass Robert J. , Klainer Albert S. , Hamburger Morton
Published: January 2020

The serial morphological changes of
Staphylococcus aureus
after exposure to lysostaphin (one unit per ml) in hypertonic culture medium containing 5% NaCl were studied over an 8-hr period at 37 C.
S. aureus
stained gram-negative almost instantaneously upon contact with lysostaphin. Many cells were lysed, but wall-defective staphylococci with corrugated surface were detected as early as 5 min by use of scanning-beam electron microscopy. Wall-defective staphylococci enlarged progressively with increasing periods of incubation. The absorbance of the lysostaphin-treated culture decreased significantly by 2 min, remained stationary for 2 hr, and then progressively increased. The number of vegetative colony-forming units decreased progressively with increasing periods of incubation, whereas the number of L-colony-forming units remained the same.
Evidence for a Bilayer Structure in Gram-Negative Lipopolysaccharide: Relationship to Toxicity
Authors: Shands J. W.
Published: January 2020

Additional evidence for a bilayer structure in isolated gram-negative lipopolysaccharide (LPS) is demonstrated. Both heating in an electron beam and cleavage of ester-bound fatty acids by alkali were found to split the bilayer structure of LPS into apparent monolayers. The altered structure of LPS resulting from alkaline hydrolysis may explain some of the altered biological activities possessed by the hydrolyzed product.
Distribution of Lysosomal Enzymes, Cationic Proteins, and Bactericidal Substances in Subcellular Fractions of Human Polymorphonuclear Leukocytes
Authors: Welsh I. R. H. , Spitznagel J. K.
Published: January 2020

Separation of homogenates of human polymorphonuclear leukocytes (PMN) into different fractions by sedimentation in centrifugal fields that ranged from 126 ×
g
to 50,000 ×
g
resulted in a differential distribution of the lysosomal enzymes. Peroxidase, lysozyme, beta-glucuronidase, and acid phosphatase activity were separated from each other. This demonstrates that the lysosomes of human PMN comprise at least three and possibly four physically and chemically different cytoplasmic particles. Proteins which are more cationic than lysozyme and which may be analogous to cationic lysosomal protein of rabbit PMN were associated with lysozyme and beta-glucuronidase rich granules. Antibacterial activity was present in four of the five cell fractions which this work produced. These results are significant because they differ from those obtained with rabbits and because they directly influence future experimental design and interpretation, in attempts to analyze antibacterial, scavenging, and inflammatory capacities of human PMN. Since lysosomes differ physically, biochemically, and morphologically, they may well differ with respect to their function in the PMN.
Mechanisms and Genetics of Resistance to Sodium Lauryl Sulfate in Strains of
Shigella
and
Escherichia coli
Authors: Corwin L. M. , Rothman Sara W. , Kim Rosalind , Talevi L. A.
Published: January 2020

The sensitivity to sodium lauryl sulfate (SLS) of
Shigella flexneri
and
Escherichia coli
is determined by at least three genes. One site is located near the lactose operon, and two loci are cotransducible with the arabinose operon. Calcium ions protect against SLS lysis. One gene is concerned with the relative ability of the bacterium to retain calcium against such chelating agents as ethylenediaminetetraacetic acid or phosphate buffer. This was first observed in a mutation from virulence to avirulence in
S. flexneri
with a concomitant loss of ability to penetrate the intestinal epithelium. The avirulent strain is far less sensitive to lysis by SLS in the presence of phosphate buffer than its virulent parent. The avirulent strain is also less sensitive to lysozyme and ethylenediaminetetraacetic acid.
E. coli
K-12 is much more sensitive to SLS than both of these
Shigella
strains.
An E. coli-S. flexneri
hybrid, which is unable to survive well in the gut and thus only produces an abortive infection, has inherited this extreme sensitivity to SLS.
Immunosuppression with
cis
-Platinum (II) Diammino-dichloride: Effect on Antibody Plaque-Forming Spleen Cells
Authors: Khan Amanullah , Hill Joseph M.
Published: January 2020

A new antitumor compound,
cis
-platinum (II) diamminodichloride, suppressed antibody plaque-forming cells in C
3
H mice when given in a single dose of 10 mg per kg of body weight within 2 days before or after sensitization.
Inhibition of
Pseudomonas aeruginosa
by Hyperbaric Oxygen I. Sulfonamide Activity Enhancement and Reversal
Authors: Pakman Leonard M.
Published: January 2020

To elucidate an explanation for in vitro sulfonamide enhancement by high-pressure oxygen (HPO) and the reported absence of enhancement with in vivo therapy,
Pseudomonas aeruginosa
cultures were exposed to selected antifolate antimicrobials in the presence of 1.87 atm absolute of O
2
and compared with non-HPO treated controls. Under these conditions, HPO alone retarded growth. Trimethoprim, a non-sulfonamide which inhibits dihydrofolate reductase, was not bactericidal, nor did HPO enhance existent bacteriostatic activity. The sulfonamide, sulfisozazole, was not bactericidal, but HPO enhanced bacteriostatic activity twofold; bacteriostasis was mitigated in HPO-treated and control cultures by
p
-aminobenzoate but not by a mixture of compounds involved in folate-mediated “1-C” biosynthesis. Mafenide, a unique sulfonamide, at high concentrations with HPO, was synergistically bactericidal; non-HPO-treated cultures were bacteriostatically inhibited. Bacteriostatic activity of lower mafenide concentrations was also enhanced at least twofold by HPO. These inhibitory effects of mafenide, acting with or without HPO, were mitigated by the above mixture, but not by
p
-aminobenzoate. This may explain the lack of in vivo HPO-mafenide enhancement in burn-wound sepsis where exudates would contain such a mixture. Lastly, HPO itself was largely bactericidal at 2.87 atm absolute of O
2
. This was reversed to various degrees by the above mixture, or its components, or by folic, folinic, or
p
-aminobenzoic acids. These in vitro interactions suggest HPO per se may act at the same site as some sulfonamides to inhibit folate synthesis (not primarily at the dihydrofolate reductase level), or coenzyme functions of folate, or both.
Cellular Immunity In Vitro: Migration Inhibition and Phagocytosis
Authors: Neta Ruth , Salvin S. B.
Published: January 2020

The activity of peritoneal exudate cells from candidin-sensitive and normal guinea pigs with and without antigen was studied in vitro with the aid of time-lapse, phase-contrast cinemicrography. Macrophages from normal animals migrate well on an agar medium and readily phagocytose Merthiolate-killed
Candida albicans
cells. A reduction in the migration of peritoneal macrophages from guinea pigs sensitized to
C. albicans
during the first 24 hr after exposure to antigen was accompanied by a decrease in phagocytosis of
C. albicans
cells. The macrophages were viable but comparatively immotile. Since the sensitized macrophages came from resistant donors, it is possible that the initial stage of cellular immunity to
C. albicans
is associated with a reduced activity of the phagocytic macrophages, apparently to limit spread of the pathogen from the infected area.
Response of human lymphocytes to measles virus after natural infection
Authors: Greenstein J I , McFarland H F
Published: January 2020

The lymphoproliferative response to measles, mumps, and vaccinia virus-infected monolayers measured in seropositive adults by thymidine incorporation demonstrated that only 5% of individuals responded well to measles virus (stimulation index, greater than 5). Possible explanations for this occurrence include a lack of sensitization, active suppression, or failure in long-term stimulation. To distinguish among these possibilities, we studied the responses to measles virus in 22 immunocompetent individuals during early convalescence from natural measles infection. Substantial responses occurred (stimulation index, 7.03), particularly in a smaller group which included those individuals with milder cases of the disease. The level of responsiveness declined over a period of weeks. Responder and nonresponder cell mixing showed no active cellular suppression. These studies indicate that the low responses to measles virus found in late convalescence represent a lack of prolonged stimulation of the cell population measured in this assay.
Biological properties of rabbit antibodies to a surface antigen of Rickettsia rickettsii
Authors: Anacker R L , Philip R N , Casper E , Todd W J , Mann R E , Johnston M R , Nauck C J
Published: January 2020

Because of the potential significance of external components of the obligate intracellular parasite Rickettsia rickettsii in host-parasite interactions, we have begun the first phase of a study to isolate and characterize surface antigens of this organism. An antiserum to a rickettsial surface component was obtained from rabbits inoculated with immune precipitates prepared by crossed immunoelectrophoresis of Triton X-100 extracts of R. rickettsii strain R. This antiserum (i) protected guinea pigs inoculated with 10,000 guinea pig 50% infectious doses of R. rickettsii against fever, (ii) prevented death of mice challenged with 2 50% lethal doses of R. rickettsii, and (iii) reacted in the microimmunofluorescence test with 9 of 13 spotted fever group serotypes tested. The location of this antigen on the rickettsial surface was demonstrated by immunoelectron microscopy with ferritin-labeled antibodies.
Culture Supernatants of
Campylobacter jejuni
Induce a Secretory Response in Jejunal Segments of Adult Rats
Authors: Fernández Heriberto , Neto Ulysses Fagundes , Fernandes Fernando , de Almeida Pedra Marly , Trabulsi Luiz R.
Published: January 2020

Culture supernatants of four
Campylobacter jejuni
strains induced a net sodium secretory flux (plasma-lumen) and an impaired glucose transport in perfused jejunal segments of adult rats in vivo.
Role of antibody and enterobactin in controlling growth of Escherichia coli in human milk and acquisition of lactoferrin- and transferrin-bound iron by Escherichia coli
Authors: Brock J H , Pickering M G , McDowall M C , Deacon A G
Published: January 2020

Growth of Escherichia coli NCTC 8623 in human milk was slow during the first 10 h of incubation, but this bacteriostatic effect had disappeared by 24 h. The bacteriostatic phase could be abolished by adding sufficient iron to saturate the lactoferrin in human milk, and also by adding supernatant from a 24-h milk culture or by adding enterobactin, an enterobacterial iron chelator. Growth in the presence of enterobactin was even more rapid than in the presence of excess iron. Partial loss of bacteriostatic activity could be achieved by absorbing the milk with bacterial antigens, but no clear correlation with removal of antibodies to O, K, or H antigens was apparent. When E. coli was grown in human serum trace-labeled with 59Fe, the organisms acquired iron from transferrin during growth. Cultivation of E. coli in a minimal medium supplemented with transferrin or lactoferrin doubly labeled with 125I and 59Fe showed that iron acquisition occurred without either assimilation or degradation of the iron-binding proteins.
Effect of indomethacin on the incidence of experimental Escherichia coli pyelonephritis
Authors: Glauser M P , Francioli P B , Bille J , Bonard M , Meylan P
Published: January 2020

In the presence of temporary obstruction (20 h), ascending Escherichia coli urinary infection leads to irreversible acute exudative pyelonephritis (AEP) in rats. The purpose of the present study was to investigate the early inflammatory events which take place in response to the presence of bacteria in the kidney parenchyma and lead to the development of AEP. Rats were given indomethacin before and during the obstructive phase of kidney infection and were sacrificed at different times thereafter. Although renal infection (as defined by bacterial counts) was equally frequent (76%) and severe in indomethacin-treated and control rats sacrificed at the end of the obstructive period, it was found that the incidence of AEP (as defined by the inflammatory response of the kidney elicited by bacteria) 2 days after removal of the obstruction was significantly reduced from 74% in controls given water to 48% in indomethacin-treated animals (P = 0.02). Rat kidneys without AEP had bacterial counts of 10(2)/g. Since indomethacin apparently had no direct antibacterial activity against E. coli and no effect on urine osmolalities, it is likely that the reduction in the incidence of AEP and the concomitant eradication of bacteria after removal of the obstruction was due to an effect of indomethacin that is related to the renal response to infection. This was possibly due to decreased inflammation, as indicated by the fact that when pyelonephritis developed in indomethacin-treated rats it was less severe than in controls. These results suggest that if inflammation can be mitigated when bacteria are present in the kidney during obstruction, the bacteria may be cleared spontaneously once the normal urinary flow is restored.
Acquisition of alpha 1-Antitrypsin by a pathogenic strain of Trichomonas vaginalis
Authors: Peterson K M , Alderete J F
Published: January 2020

The interaction of alpha 1-Antitrypsin, the major serine protease inhibitor in plasma, with pathogenic Trichomonas vaginalis and the acquisition by trichomonads of this host protein from normal human plasma were investigated. alpha 1-Antitrypsin acquired by intact parasites could not be removed by repeated washings in phosphate-buffered saline. Saturation kinetics were observed after incubation of glutaraldehyde-fixed organisms with 125I-labeled alpha 1-antitrypsin. Evidence suggesting that specific parasite membrane sites were responsible for trichomonal acquisition of alpha 1-antitrypsin was obtained through competitive binding experiments using purified preparations of homologous versus heterologous plasma proteins. No evidence of degradation of bound antitrypsin by live parasites was observed. The avid binding of alpha 1-antitrypsin by pathogenic T. vaginalis after incubation in normal human plasma was demonstrated by using sensitive electrophoretic and immunodetection techniques. Radioimmunoprecipitation of intrinsically labeled, detergent-solubilized extracts of T. vaginalis incubated with monospecific antisera against alpha 1-antitrypsin and other human plasma proteins revealed the inability of parasites to biosynthesize any substance cross-reactive with host plasma proteins. Finally, T. vaginalis organisms pretreated with alpha 1-antitrypsin inhibited trypsin caseinase activity in an in vitro assay. The implications of these observations are discussed.
Broad geographical distribution of a cytotoxin gene mediating beta-hemolysis and bacteriocin activity among Streptococcus faecalis strains
Authors: LeBlanc D J , Lee L N , Clewell D B , Behnke D
Published: January 2020

Conjugative hemolysin-bacteriocin plasmids were isolated from Streptococcus faecalis var. zymogenes strains of diverse geographical origins. Cloned DNA fragments containing the hemolysin-bacteriocin gene(s) from one of these plasmids (pAD1) hybridized to two EcoRI fragments of identical size from each of the five plasmids examined. Results of hybridization experiments in which total plasmid DNA was used suggested that the plasmids shared extensive homology. Two of the plasmids, pAD1 from strain DS16 (Ann Arbor, Mich.) and pAM gamma 1 from strain DS5 (Miami, Fla.), were 100% homologous and had identical EcoRI restriction patterns (eight fragments each). There was no detectable homology between the plasmid-mediated hemolysin determinants of S. faecalis and DNA from other beta-hemolytic streptococci (Lancefield groups A, B, F, or H).
Procholeragenoid: a safe and effective antigen for oral immunization against experimental cholera
Authors: Pierce N F , Cray W C , Sacci J B , Craig J P , Germanier R , Fürer E
Published: January 2020

The immunogenicity and safety of procholeragenoid, a minimally toxic, heat-induced aggregate of cholera toxin (CT), were studied in enterically immunized rats and dogs. Although 99% less toxic than CT, procholeragenoid was only slightly less efficient in causing jejunal anti-CT responses in rats; in contrast, choleragenoid, the nontoxic B subunit pentamer of CT, was much less effective. The immunogenicity of procholeragenoid was due almost entirely to its large-molecular-weight components (MW = 10(6) to 10(7)) and was markedly reduced by preincubation with GM1 ganglioside or treatment with Formalin to eliminate residual toxicity. These findings suggest that molecular aggregation, binding to GM1 receptors on cell membranes, and stimulation of cellular adenylate cyclase each contributed to the effectiveness of procholeragenoid as a mucosal immunogen. In dogs, oral immunization with five 500-micrograms doses of procholeragenoid evoked vigorous anti-CT responses in jejunal mucosa without causing significant diarrhea. When subsequently challenged with virulent Vibrio cholerae, immunized dogs showed 83% protection against the development of severe or lethal diarrhea compared with non-immunized controls. These results confirm a protective role for mucosal antitoxin in experimental cholera and show that procholeragenoid is both safe and effective as an oral immunogen. Procholeragenoid, combined with other antigens of V. cholerae, may constitute a simple, safe, and effective oral vaccine for cholera.
Mechanism of immunosuppression in leprosy: presence of suppressor factor(s) from macrophages of lepromatous patients
Authors: Salgame P R , Mahadevan P R , Antia N H
Published: January 2020

Human peripheral blood mononuclear cell proliferation induced by Mycobacterium leprae could be inhibited by the suppressor factor in the lysate of the macrophages of lepromatous leprosy patients. Macrophages from normal subjects and tuberculoid patients did not show production of a suppressor factor. Inhibition occurred only when the factor was present in the initial stages of lymphocyte culture. The factor is heat stable and nondialyzable. Proliferation induced by some mycobacteria and concanavalin A could also be blocked by the factor. Interestingly, blastogenic response by a few other antigens and phytohemagglutinin could not be inhibited by the suppressor factor. Mononuclear cells pretreated with such lysate from lepromatous macrophages for 24 h could induce suppressive activity in the cells in vitro in an autologous system. Treatment of these cells with carbonyl iron after the induction phase, to remove phagocytic cells, did not abolish their suppressive activity. The lepromatous macrophage lysate also generated suppressive activity in a T-lymphocyte-enriched population of normal subjects. These studies are interpreted to indicate that immunosuppression in lepromatous patients is produced by both macrophages and T lymphocytes. The exact phase in which either of these cells acts as a suppressor may be different. Specific suppression by macrophages to M. leprae can be an early event, and nonspecific suppression by T lymphocytes may be a later event in the course of lepromatous leprosy.
Course of infection and development of immunity in experimental infection of mice with Listeria serotypes
Authors: von Koenig C H , Heymer B , Hof H , Finger H
Published: January 2020

NMRI mice were experimentally infected with Listeria monocytogenes serotypes 1/2b, 3a, 4b, and 4d and Listeria innocua serotype 6b by different means. The course of infection was monitored, using bacteriological and histological methods. The following typical features of experimental infection with the various L. monocytogenes and L. innocua serotypes were observed. (i) On the basis of the mean lethal dose, L. monocytogenes 4b, 4d, and 1/2b proved to be mouse pathogenic, although to different degrees, L. monocytogenes 3a and L. innocua can be regarded as nonpathogenic for NMRI mice. The virulence of L. monocytogenes serotype 4d was increased 1,000-fold after adaptation to mice. (ii) Primary infection with any serotype of L. monocytogenes or L. innocua resulted in protection against a lethal challenge with the most virulent serotype, 4b. This protective immunity could be transferred by spleen cells. Compared with the duration of immunity achieved by infection with L. monocytogenes serotype 4b, the protection induced by infection with L. innocua was short lived and dose dependent. The data obtained also suggest that immunity after experimental infection with any serotype of L. monocytogenes or L. innocua is produced only when the animal host is filled with bacteria. (iii) The distribution of the germs in the internal organs of the mouse shortly after infection was dependent on the route of infection rather than on the serotype used. (iv) The main difference among the Listeria serotypes tested was their ability to multiply within the host and to induce a granulomatous inflammation. The results indicate that mouse pathogenicity and virulence of Listeria spp. cannot be defined only by the capacity of the bacteria to infect or kill conventional mice. Such a definition should include an analysis of the immune system of the host, a kinetic study of experimental infection, and a histomorphological evaluation of the lesions induced.
Host defenses in murine malaria: characteristics of protracted states of immunity to Plasmodium berghei
Authors: Murphy J R
Published: January 2020

Vaccination of 8-week-old B6D2 (C57BL/6 x DBA/2) mice with Formalin-killed asexual erythrocytic stages of Plasmodium berghei provided a capacity to survive challenge with lethal P. berghei for more than 542 days. Although long-lived, this immunity did not provide a capacity to immediately neutralize parasites in a challenge; significant levels of erythrocytic infection occurred transiently after each challenge. The quality of long-lived immunity was not enhanced by an injection of live parasites 2 weeks after the last injection of killed antigen. The results suggest that it will be difficult to design a vaccination regimen which will provide long-lived protection from overt erythrocytic-stage infection.
Murine encephalitozoonosis model for studying the host-parasite relationship of a chronic infection
Authors: Schmidt E C , Shadduck J A
Published: January 2020

Encephalitozoon cuniculi caused chronic nonlethal infections in euthymic BALB/cAnN mice, whereas athymic BALB/cAnN-nu mice died from infection. Specific, anamnestic, transferable, and acquired responses against E. cuniculi were expressed by infected euthymic mice. Resistance to lethal disease appears to be T-cell dependent. Immune serum failed to protect infected athymic mice, whereas the transfer of T-enriched spleen cells from E. cuniculi-sensitized euthymic donors prevented lethal E. cuniculi infections in athymic mice. These findings indicate that murine encephalitozoonosis may be an excellent system for studies of a chronic infection in an immunologically well-characterized host.
Sequence of the amino-terminal part of enterotoxin from Clostridium perfringens type A: identification of points of trypsin activation
Authors: Richardson M , Granum P E
Published: January 2020

The sequence of the first 66 amino acids of the amino-terminal part of the enterotoxin from Clostridium perfringens type A is presented. The trypsin activation of the enterotoxin involves hydrolysis of Lys15-Glu16 and Lys25-Thr26 bonds. The N-terminal sequence of the trypsin-activated enterotoxin has limited homology with the sequence of the N-terminal region of the cholera toxin B subunit.
Serology of oral Actinobacillus actinomycetemcomitans and serotype distribution in human periodontal disease
Authors: Zambon J J , Slots J , Genco R J
Published: January 2020

Actinobacillus actinomycetemcomitans from the human oral cavity was serologically characterized with rabbit antisera to the type strain NCTC 9710; a number of reference strains, including Y4, ATCC 29522, ATCC 29523, ATCC 29524, NCTC 9709; and our own isolates representative of each of 10 biotypes. Using immunoabsorbed antisera, we identified three distinct serotypes by immunodiffusion and indirect immunofluorescence. Serotype a was represented by ATCC 29523 and SUNYaB 75; serotype b was represented by ATCC 29522 and Y4; and serotype c was represented by NCTC 9710 and SUNYaB 67. Indirect immunofluorescence revealed no reaction between the three A. actinomycetemcomitans serotype-specific antisera and 62 strains representing 23 major oral bacterial species. Distinct from the serotype antigens were at least one A. actinomycetemcomitans species common antigen and an antigen shared with other Actinobacillus species, Haemophilus aphrophilus, and Haemophilus paraphrophilus. All serotype a A. actinomycetemcomitans strains failed to ferment xylose, whereas all serotype b organisms fermented xylose. Serotype c included xylose-positive as well as xylose-negative strains. A total of 301 isolates of A. actinomycetemcomitans from the oral cavity of 74 subjects were serologically categorized by indirect immunofluorescence with serotype-specific rabbit antisera. Each patient harbored only one serotype of A. actinomycetemcomitans. Fourteen healthy subjects, five diabetics, and seventeen adult periodontitis patients exhibited serotypes a and b in approximately equal frequency, whereas serotype c was found less frequently. In contrast, in 29 localized juvenile periodontitis patients, the incidence of serotype b was approximately two times higher than that of serotypes a or c, suggesting a particularly high periodontopathic potential of A. actinomycetemcomitans serotype b strains. In subjects infected with A. actinomycetemcomitans, serum antibodies were detected to the serotype antigens, indicating that these antigens may play a role in the pathogenesis of periodontal disease.
Pathogenesis of bloodstream invasion with Haemophilus influenzae type b
Authors: Rubin L G , Moxon E R
Published: January 2020

Possible route(s) by which encapsulated bacteria invade the blood from the nasopharynx include (i) the direct invasion of submucosal blood vessels and (ii) clearance via lymphatics to regional nodes followed by bloodstream invasion. These possibilities were investigated in rats after intranasal inoculation with 10(5) Haemophilus influenzae type b. Within 24 h of inoculation, 10 of 42 rats with sterile blood cultures had similar numbers of H. influenzae b recovered from both cervical (local) and periiliac (distant) lymph nodes, which suggested early bacteremic spread. When virtually continuous blood cultures were obtained for 30 min after inoculation with 10(8) H. influenzae b, early transient bacteremia was documented in four of eight rats. Also, we found no significant difference in bacteremia among rats whose cervical lymph nodes had been removed surgically compared with sham-operated rats. These findings favor the hypothesis of a rapid, perhaps direct invasion of pharyngeal blood vessels as an initial determinant of the systemic spread of H. influenzae b.
Local immune response to repeated topical antigen application in the simian labial mucosa
Authors: Nair P N , Schroeder H E
Published: January 2020

Minor salivary glands of the oral mucosa in healthy monkeys (Macaca fascicularis and Macaca mulatta) contain organized structural units suitable for recognizing and processing antigens. A previous study of M. fascicularis monkeys provided experimental evidence of retrograde access of oral antigens deep into the minor salivary glands. The present study aimed at exploring the possible immune response of simian oral mucosa to repeated topical application of a chemically defined antigenic solution at the labial and gut mucosa. Ten female M. fascicularis animals were challenged topically at the lower lip mucosa at weekly intervals for a variable period of 4 to 8 weeks with a solution consisting of horseradish peroxidase, ferritin, and special India ink. Transmission electron microscopic examination of immunohistochemically treated sections of the labial glands revealed the presence of plasma cells containing specific anti-horseradish peroxidase antibody. These cells resided in the interacinar regions. Enteric and gut priming with the same antigen in four other monkeys, bypassing the oral mucosa, failed to reveal the presence of horseradish peroxidase-positive plasma cells in the labial mucosa of any of the four animals, although in one animal such cells could be identified in a mesenteric lymph node. This is suggestive of the existence, at least in primates, of a local immune response of the oral mucosa independent of systemic involvement.
Analysis of antigenic determinants in cholera enterotoxin and heat-labile enterotoxins from human and porcine enterotoxigenic Escherichia coli
Authors: Takeda Y , Honda T , Sima H , Tsuji T , Miwatani T
Published: January 2020

Antigenic determinants of cholera enterotoxin (CT) and heat-labile enterotoxin from a human strain (LTh) and a porcine strain (LTp) were analyzed by Ouchterlony double-gel diffusion test against anti-CT, anti-LTh, and anti-LTp, which were treated by immunoaffinity column chromatography. The results showed the existence of the following antigenic determinants: (i) antigenic determinants unique to CT, LTh, and LTp, respectively; (ii) an antigenic determinant common to CT, LTh, and LTp; (iii) an antigenic determinant common to CT and LTh, but not LTp; and (iv) an antigenic determinant common to LTh and LTp, but not CT. On the basis of these results, an antigenic scheme for CT, LTh, and LTp is proposed.
Further comparisons of endogenous pyrogens and leukocytic endogenous mediators
Authors: Kampschmidt R F , Upchurch H F , Worthington M L
Published: January 2020

It was recently shown (Murphy et al., Infect. Immun. 34:177-183), that rabbit macrophages produce two biochemically and immunologically distinct endogenous pyrogens. One of these has or copurifies with substances having a molecular weight of 13,000 and a pI of 7.3. This protein was produced by blood monocytes or inflammatory cells elicited in 16-h rabbit peritoneal exudates. These acute peritoneal exudates were produced by the intraperitoneal injection of large volumes of saline containing shellfish glycogen. When the leukocytes in these exudates were washed and incubated at 37 degrees C in saline, they released an endogenous pyrogen. The injection of this pyrogen into rabbits, rats, or mice caused the biological manifestations which have been attributed to leukocytic endogenous mediator. These effects were increases in blood neutrophils, the lowering of plasma iron and zinc levels, and the increased synthesis of the acute-phase proteins. The other rabbit endogenous pyrogen seems to be a family of proteins with isoelectric points between 4.5 and 5.0. These proteins are produced by macrophages in the lung, liver, or in chronic peritoneal exudates. In these experiments, the lower-isoelectric-point endogenous pyrogens were produced by macrophages from the peritoneal cavity of rabbits that had been injected 4 days earlier with 50 ml of light mineral oil. These rabbit pyrogens were found to have leukocytic endogenous mediator activity in mice but to be completely inactive in rats. When injected into rabbits, these proteins produced fever, lowered plasma iron, increased blood neutrophils, but failed to elevate plasma fibrinogen.
Autoimmunity in Aleutian disease: contribution of antiviral and anti-DNA antibody to hypergammaglobulinemia
Authors: Hahn E C , Hahn P S
Published: January 2020

The contributions to Aleutian disease gammopathy of specific antiviral antibody and an autoimmune component, anti-DNA antibody, were studied with pastel ranch mink naturally infected with Aleutian disease virus. Specific antibody activities were determined by countercurrent immunoelectrophoresis and radioimmune assay, respectively. Gamma globulin levels (percent gamma) were determined by serum electrophoresis. Within an infected mink population, it was possible to predict the level of gammopathy from measurement of the two antibody levels. For the mink serum samples used, there was better correlation between anti-DNA antibody levels and total serum immunoglobulin than between anti-Aleutian disease virus antibody titers and percent gamma. With serum samples taken over a 2-week interval, significant increases were measured in anti-DNA antibody and percent gamma. Increases in anti-Aleutian disease virus titers during this period were not significant. The results suggest that the continuing increases in serum immunoglobulin in Aleutian disease virus-infected mink are due to both a specific antiviral response and an autoimmune response, as reflected in generation of anti-DNA antibody.
Effects of substitution on polyglycerol phosphate-specific antibody binding to lipoteichoic acids
Authors: Kessler R E , Thivierge B H
Published: January 2020

The influence of D-alanine and carbohydrate substitution of lipoteichoic acids (LTAs) on the binding of antibody directed to the polyglycerol phosphate (PGP) portion was found to be at least partially dependent upon the mode of presentation of the antigen. There were greater differences in binding of anti-PGP immunoglobulins to substituted and unsubstituted LTAs in solution (micellar presentation) than when the same LTAs were adsorbed to the erythrocyte surface, which suggests that there is greater hindrance of access to the PGP chain, possibly as a result of closer packing, in a vesicle or micelle than when LTA is bound to a membrane surface. Although the difference in binding of anti-PGP immunoglobulins was nearly 20-fold between unsubstituted and highly substituted LTAs, rocket heights by rocket immunoelectrophoresis were only 4-fold different for the most highly substituted LTA and unsubstituted LTA. However, unsubstituted LTA clearly bound more immunoglobulin molecules than was reflected in the rocket height alone, since the resulting immunoprecipitates were much more prominent or intense, both before and after staining, than those of highly substituted LTAs. Differences between lightly and moderately substituted LTAs were less than twofold, indicating that under most circumstances estimates of LTA concentrations in samples where the composition is unknown will be within approximately twofold of the estimate.
Production of cholera toxin-like toxin by Vibrio mimicus and non-O1 Vibrio cholerae: batch culture conditions for optimum yields and isolation of hypertoxigenic lincomycin-resistant mutants
Authors: Spira W M , Fedorka-Cray P J
Published: January 2020

Vibrio mimicus 61892, isolated in 1977 from a case of watery diarrhea in Bangladesh, produces an enterotoxin which possesses activity in Y-1 mouse adrenal cells and in rabbit ileal loops which is identical to the prototype cholera toxin (CT) produced by Vibrio cholerae 569B. The neutralization of the adrenal cell activity of 61892 toxin and 569B CT by homologous and heterologous antisera generates parallel titration curves which show complete neutralization in all cases. Paired titrations in the ganglioside GM1 enzyme-linked immunosorbent assay (using either CT or Escherichia coli heat-labile toxin antitoxin) of both toxins indicates that 61892 toxin is antigenically indistinguishable from 569B CT. The specific activity of the two toxins in the rabbit ileal loop is virtually identical. Batch culture production of CT-like toxin and CT by isolates of V. mimicus and different biotypes of V. cholerae was found to be highest in shake flask cultures of Casamino Acids-yeast extract broth grown at 27 degrees C with vigorous aeration. Incorporation of lincomycin into the growth medium at a concentration of 50 micrograms/ml increased yields from wild-type strains. Dramatically higher yields were obtained when a spontaneous resistance mutant of strain 61892 was grown in the presence of 200 to 300 micrograms of lincomycin per ml. Under these conditions, yields of CT-like toxin were increased by 300- to 500-fold, and the highest yields reached more than 100 micrograms/ml after 44 h of culture. This is substantially higher than that reported in the literature for CT production by any strain of V. cholerae, including hypertoxigenic strain 569B.
Study of heart-reactive antibody in antisera and hybridoma culture fluids against group A streptococci
Authors: Cunningham M W , Russell S M
Published: January 2020

Heart-reactive antibody is known to be produced in animals immunized with group A streptococci. However, the detection and quantitation of these antibodies and their respective antigens have been limited to immunofluorescence or immunoprecipitation tests. In this study a more sensitive technique was used to detect heart-reactive antibody, i.e., the enzyme-linked immunosorbent assay (ELISA). Countercurrent immunoelectrophoresis and agar gel immunodiffusion were also used to confirm the reactivity of rabbit antisera to group A streptococci and human heart tissue. Human heart tissue antigen was prepared by Triton X-100 extraction, and checkerboard titrations of heart tissue antigen versus streptococcal antisera revealed optimal concentrations of each for the ELISA. When immune rabbit serum was reacted with heart tissue antigen, a 5- to 10-fold increase was observed over the reactions of preimmune sera controls. Streptococcal antiserum was found to have a six- to eightfold greater reactivity with heart tissue antigen than with similar concentrations of kidney and skeletal muscle antigens. Heart tissue antigen was partially purified by DEAE-cellulose chromatography, and quantities of greater than 10 to 100 ng (dry weight) were shown to react with streptococcal antisera in the ELISA. Heart, kidney, and skeletal muscle antigens were subjected to electrophoresis in polyacrylamide gel slabs and blotted onto a nitrocellulose filter. These filter blots reacted with streptococcal antisera and confirmed the tissue antigen specificity observed with the ELISA. In addition, the ELISA was found to be an effective method for the detection of heart-reactive antibodies produced by murine hybridomas that were producing antibody to group A streptococci.
In vitro and in vivo complementation of Streptococcus mutans mutants defective in adherence
Authors: Larrimore S , Murchison H , Shiota T , Michalek S M , Curtiss R
Published: January 2020

Previous studies have shown that adherence-defective mutants of Streptococcus mutans PS14, serotype c, can be grouped into several different phenotypic groups. In this study a method was developed to test for complementation between pairs of nonadhering mutants which possess different genotypic defects. Mutant strains UAB95 and a spectinomycin-resistant derivative of UAB95 (UAB516) were found to exhibit increased levels of adherence when grown together with UAB230 in media containing sucrose as compared to the adherence of each strain grown separately. An increase in caries was also observed in gnotobiotic rats mixedly infected with the two mutants as compared to either strain alone. Tests revealed that UAB95 produced more water-insoluble glucan than its parent strain but had a defect in glucan binding. UAB230 was found to produce levels of a defective glucan that could not be bound by mutant or wild-type cells. Our results suggest that UAB95 produces a water-insoluble glucan which is bound by UAB230, thus allowing complementation for adherence and caries production.
Host resistance of mice to Naegleria fowleri infections
Authors: Reilly M F , White K L , Bradley S G
Published: January 2020

Naegleria fowleri is an etiological agent of primary amoebic meningoencephalitis in humans and laboratory animals. The determinative factors in host resistance of mice to N. fowleri infections have not been fully characterized. Male or female B6C3F1 mice stimulated by intraperitoneal administration of 10(6) amoebae of N. fowleri nN68 per mouse produced agglutinating activity and markedly elevated levels of serum and immunoglobulins M and G. Despite a marked humoral response, protective immunity was increased only marginally by active immunization. Host resistance was not impaired by prior treatment with 350 rads of 60Co radiation or 200 mg of cyclophosphamide per kg or by concurrent daily treatment with 30 mg of cyclophosphamide per kg for 14 days. Moreover, host resistance was not impaired by daily treatment with 4 mg of diethylstilbestrol per kg for 14 days, with challenge on day 2 of drug exposure or 24 h after the last drug treatment. Mice depleted of hemolytic complement by cobra venom factor were more susceptible to N. fowleri infection than were untreated mice.
X-linked resistance of mice to high doses of herpes simplex virus type 2 correlates with early interferon production
Authors: Pedersen E B , Haahr S , Mogensen S C
Published: January 2020

Mice inoculated intraperitoneally with herpes simplex virus type 2 develop focal necrotizing hepatitis and eventually die from ascending myelitis and encephalitis. The genetics of resistance to the infection were analyzed in crosses between resistant C57BL/10 mice and susceptible BALB/c mice. It was shown that the resistance of C57BL/10 mice to hepatitis induction was influenced by an X-linked dominant gene as previously shown for the GR mouse strain. The course of infection in the liver pointed to early, natural defense mechanisms as being responsible for the difference between the mouse strains, whereas the clearance of virus from the liver, probably mediated by specific immunity, was exerted at the same time and with equal efficiency for all groups of mice. In mortality experiments, resistance was shown to be an autointerference phenomenon in that a considerable number of C57BL/10 mice survived an intraperitoneal injection of 10(6) PFU, whereas all mice were killed by 10(5) PFU. This resistance of C57BL/10 mice to high doses of HSV-2 was retrieved in all groups of F1 mice in crosses between C57BL/10 and BALB/c mice except the (BALB/c female X C57 male) male group, in which the mice receive the X chromosome from the susceptible BALB/c female. Thus, the autointerference phenomenon also seems to be influenced by loci on the X chromosome. A similar pattern of inheritance was observed when early interferon induction (4 to 5 h after infection) in response to HSV-2 was measured. The possible relevance of this early interferon response in conjunction with other potential natural defense mechanisms is discussed.
Insoluble glucan synthesis by Streptococcus mutans serotype c strains
Authors: Kuramitsu H K , Wondrack L
Published: January 2020

Both dextransucrase and mutansynthetase activities have been purified from the culture fluids of Streptococcus mutans GS-5 (serotype c). Although homogeneous dextransucrase preparations normally synthesize little insoluble glucan, essentially all of the glucan synthesized by this enzyme in the presence of 1.5 M (NH4)2SO4 was water insoluble. Linkage analysis of the insoluble glucans indicated that the presence of NH4+ increased the portion of alpha-1,3-glucose linkages relative to alpha-1,6-glucose units in the product. Chromatofocusing of aggregated glucosyltransferase fractions synthesizing predominantly insoluble glucan yielded primarily dextransucrase activity separable from relatively low levels of mutansynthetase activity. The latter enzyme was detected only in 18-h assays and synthesized primer-dependent insoluble glucan, which was decreased in the presence of NH4+. In the absence of primer dextran T10, the addition of dextransucrase also stimulated insoluble glucan synthesis by mutansynthetase. Dextransucrase and mutansynthetase appear to be distinct enzymes, since the latter possesses a higher molecular weight (155,000 compared to 140,000), a much lower isoelectric point, and did not cross-react with antibody directed against dextransucrase. These results are discussed relative to the mechanism of insoluble glucan synthesis by S. mutans serotype c strains.
Experimental allergic neuritis-like disease in rabbits after injection with influenza vaccines mixed with gangliosides and adjuvants
Authors: Ziegler D W , Gardner J J , Warfield D T , Walls H H
Published: January 2020

An experimental allergic neuritis-like disease was induced in rabbits 3 to 8 weeks after injection with large doses of influenza vaccines mixed with gangliosides, cholesterol, and Freund complete adjuvant. The inclusion of gangliosides was essential to induce the experimental allergic neuritis-like disease. In trials with six different lots of vaccine, both swine influenza and non-swine influenza vaccines produced by four different manufacturers induced experimental allergic neuritis-like disease in 26 of 43 inoculated rabbits.
Role of macrophages in host defense against hepatic amoebiasis in hamsters
Authors: Ghadirian E , Meerovitch E , Kongshavn P A
Published: January 2020

The role of macrophages in hepatic amoebiasis in hamsters has been investigated by means of antimacrophage serum prepared in rabbits. Animals treated with normal rabbit serum or antimacrophage serum, as well as untreated controls, were inoculated intrahepatically with 10(5) axenic trophozoites of Entamoeba histolytica. In hamsters treated with antimacrophage serum before intrahepatic inoculation of amoebae, the mean weight of the metastatic foci was significantly greater than in normal rabbit serum-treated or untreated controls. Treatment of hamsters with antimacrophage serum both before and after administration of amoebae not only increased significantly the size of the abscess in the liver but also allowed dissemination of metastatic foci to other organs.
In vivo degradation of gonococcal outer membrane proteins within human leukocyte phagolysosomes
Authors: Eaton L J , Rest R F
Published: January 2020

We previously showed in vitro hydrolysis of outer membrane proteins by lysosomal proteases and purified elastase. In this study we examined the in vivo relevance of the previous studies. Outer membranes were obtained from Neisseria gonorrhoeae type 3 (strain GC7) by LiCl2 extraction. Some preparations were labeled with 125I. Phagocytizable particles were prepared by coating latex beads with outer membranes, and polymorphonuclear leukocytes were allowed to phagocytize serum-opsonized particles. After homogenization of neutrophils, phagolysosomes were recovered by flotation through sucrose. Phagolysosomes were prepared for slab gel electrophoresis immediately or incubated further at 37 degrees C to allow continued degradation of outer membrane proteins. The principal protein (protein I) and minor proteins (proteins II) of outer membranes were hydrolyzed in whole neutrophils and in isolated phagolysosomes. Proteins II were more susceptible to hydrolysis than protein I. Hydrolytic products formed were nearly identical in vivo and in vitro. We also radiolabeled the surface-exposed proteins of live gonococci. Degradation of outer membrane proteins on the intact bacteria within neutrophil and monocyte phagolysosomes was shown. This indicates that our earlier in vitro model is relevant to in vivo hydrolysis of gonococcal outer membrane proteins.