bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro tuberculin reactivity of lymphocytes from patients with tuberculous pleurisy
Authors: Fujiwara H , Okuda Y , Fukukawa T , Tsuyuguchi I
Published: January 2020

Mononuclear cells in pleural fluid from patients with tuberculous pleurisy were predominantly T cells. Responsiveness of pleural fluid T cells to purified protein derivative of tuberculin were studied by the assay of cell proliferation and production of lymphocyte mitogenic factor by the stimulation with purified protein derivative. Peripheral blood lymphocytes were also studied from patients and tuberculin-positive healthy controls. The order of responsiveness was as follows: pleural fluid lymphocytes greater than peripheral blood lymphocytes of patients without effusion = peripheral blood lymphocytes of healthy controls greater than peripheral blood lymphocytes of patients with effusion. The poor response of peripheral blood lymphocytes from pleurisy patients were recovered by the elimination of adherent cells in peripheral blood lymphocytes to the level of the response of peripheral blood lymphocytes from healthy controls. T cells purified from pleural fluid mononuclear cells responded more than those from peripheral blood. These results suggested that in the pleurisy patients purified protein derivative-reactive T cells in peripheral blood did not decrease in activity, but were depressed by suppressor cells, and further suggested that highly purified protein derivative-reactive T cells were accumulated in the pleural fluid.
Effect of trace metals on growth of Streptococcus mutans in a teflon chemostat
Authors: Aranha H , Strachan R C , Arceneaux J E , Byers B R
Published: January 2020

Correlations between the presence of certain trace metals in dental enamel or in drinking water and the incidence of human dental caries have been demonstrated; therefore, the effects of several trace metals on growth of the cariogenic organism Streptococcus mutans OMZ176 were determined. For continuous growth in a chemically defined medium (treated with Chelex-100 to lower trace metal contamination and supplemented with high-purity trace metal salts) used in a chemostat constructed of Teflon, S. mutans required input of carbon dioxide and supplementation with magnesium (126 microM) and manganese (18 to 54 microM). Addition of iron (3.6 microM) increased the level of steady-state growth by a factor of 2.8 (stimulation index [SI]); zinc at 0.4 microM nearly doubled equilibrium growth (SI = 0.9). Higher concentrations of iron and zinc (5.4 and 0.8 microM, respectively) were less stimulatory (SI values of 1.95 and 0.3, respectively). Small (but statistically significant) increases in steady-state growth were effected by cobalt (SI = 0.3 at 5.1 to 20.4 microM) and tin (SI = 0.4 at 5.1 to 10.2 microM). These data suggest nutritional requirements for these metals. Copper at a concentration of 0.16 microM was inhibitory. These results show significant effects of these metals on growth of S. mutans and may confirm epidemiological evidence suggesting a role for certain trace metals in the incidence of dental caries.
Polypeptides and functions of antigens from human coronaviruses 229E and OC43
Authors: Schmidt O W , Kenny G E
Published: January 2020

Coronaviruses possess three major size classes of polypeptides as judged by molecular weight: approximately 180,000, approximately 50,000, and approximately 23,000. Human coronaviruses 229E and OC43 possess not only three similar size classes of polypeptides but also three distinct antigens, none of which cross-react with the heterologous strain. Polypeptides separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were reacted in rocket immunoelectrophoresis with antiserum monospecific to each of the three strain-specific antigens (excised precipitin lines from crossed immunoelectrophoresis profiles were used for immunogens). Monospecific antiserum with neutralizing ability reacted with a polypeptide of 186,000 daltons for 229E and a polypeptide of 190,000 daltons for OC43. The antigen which elicited neutralizing antibody response was located at the surface, associated with the corona of the virion, glycosylated, and bound by concanavalin A. Another less prominent surface antigen was represented by size classes of 23,000 daltons for 229E and 24,000 for OC43. The core antigens of the viruses had molecular weights of 49,000 and 229E and 52,000 and OC43 virus. Thus, the molecular weights and functions of the antigens of human coronaviruses are similar to those of animal coronaviruses. The polypeptides of coronaviruses 229E and OC43 are nearly identical as judged by molecular weight, but the similar polypeptides of the two viruses represent different immunological specificities.
Experimental Escherichia coli ascending pyelonephritis in rats: changes in bacterial properties and the immune response to surface antigens
Authors: Mattsby-Baltzer I , Hanson L A , Kaijser B , Larsson P , Olling S , Svanborg-Edén C
Published: January 2020

Systemic and urinary antibody responses were examined in rats with experimental ascending pyelonephritis caused by Escherichia coli O6K13H1. During 12-month follow-up of the infections, bacterial characteristics of the urinary and renal isolates were followed: O and K antigenicity, sensitivity to the bactericidal effect of normal human serum, capacity to attach to urinary tract epithelial cells, hemolytic activity, biochemical pattern, and virulence. During the long-term infection, the urinary and renal bacterial isolates changed in O and K antigenicity, serum sensitivity, and virulence. The adhesive capacity of the bacterial isolates did not change, possibly explaining the persistence of the bacteria in the urinary tract. The serum anti-O6 antibody levels remained high during the entire 1-year observation period, especially in the rats with renal involvement. Urinary anti-O6 antibodies were also found. The serum and urinary antibodies could have played a role in bringing about the observed changes in bacterial characteristics. Antibodies to lipid A were recorded in 9 of 16 rats with pyelonephritis and renal scarring and in 1 of 9 rats not having pyelonephritis or renal bacterial growth.
Further characterization of complement resistance conferred on Escherichia coli by the plasmid genes traT of R100 and iss of ColV,I-K94
Authors: Binns M M , Mayden J , Levine R P
Published: January 2020

We have shown that the traT gene product was responsible for the complement resistance of the R100 plasmid. We compared this resistance with that specified by the iss gene of the ColV,I-K94 plasmid. The levels of resistance specified by the two genes were similar, and there was no additive effect on resistance when both genes were present together. Under conditions in which traT and iss conferred at least a 50- and 10-fold increase in survival, respectively, the consumption of C6, C7, C8, and C9 was the same for bacteria with and without the plasmid genes. This result indicated that it was the action of the terminal complex, not its formation, which was blocked by traT and iss.
Specificity of natural antibodies reactive with Streptococcus mutans in monkeys
Authors: Russell R R , Beighton D
Published: January 2020

Enzyme-linked immunosorbent assays were used to measure levels of natural antibody to defined antigens of Streptococcus mutans in sera from monkeys (Macaca fascicularis). The results suggest that most of the antibody in young monkeys that binds to whole bacteria is not specific to S. mutans.
Immunochemical and cytotoxic activities of Shigella dysenteriae 1 (shiga) and shiga-like toxins
Authors: O'Brien A D , Laveck G D
Published: January 2020

Toxins in culture supernatants and bacterial lysates of S. dysenteriae 1 and S. flexneri were quantitated by a cytotoxicity assay and a newly developed radioimmunoassay. Cytotoxin titers paralleled toxin antigen levels. Thus, variations in cytotoxicity among shigellae probably reflect differences in toxin yield rather than specific activity (cytotoxicity per microgram of toxin antigen).
Identification of a new genetic site (sac-3+) in Neisseria gonorrhoeae that affects sensitivity to normal human serum
Authors: Shafer W M , Guymon L F , Sparling P F
Published: January 2020

A previously undescribed genetic site (sac-3) affecting susceptibility of the gonococcus to normal human serum was localized on the gonococcal chromosome. The presence of the sac-3+ allele in a clinical isolate (FA889) resulted in sensitivity only to relatively high concentrations of serum (greater than or equal to 12.5%). Genetic mapping experiments demonstrated that sac-3+ was tightly linked to another genetic site (sac-1+) involved in determining susceptibility to normal human serum and to a locus (nmp-3) involved in the replacement of outer membrane protein I. The sac-1+ and sac-3+ loci resulted in phenotypically distinct levels of sensitivity to human serum. The sac-3+ serum sensitivity and sac-1+ serum sensitivity loci recombined with high frequency, resulting in serum resistance. The results show that serum sensitivity in clinical isolates may be due to different serum sensitivity loci and suggest that different antigens and immunological mechanisms could be responsible for sensitivity of different gonococcal isolates to human serum.
Immunization of cattle with a variant-specific surface antigen of Trypanosoma brucei: influence of different adjuvants
Authors: Wells P W , Emery D L , Hinson C A , Morrison W I , Murray M
Published: January 2020

Nine different adjuvants were examined for their ability to potentiate the humoral and cell-mediated immune responses of cattle to a soluble glycoprotein antigen prepared from Trypanosoma brucei. Serological responses as measured by the Farr assay were best augmented by the oil-based adjuvants and saponin. Cell-mediated immunity, as assessed by specific lymphocyte transformation in vitro, was enhanced by all oil-based adjuvants at different intervals after immunization. Results from a challenge infection of immunized cattle with the homologous clone of T. brucei and from neutralization tests indicated that protection against infection was better correlated with specific antibodies than with cell-mediated responses. From these considerations, and the absence of tissue reactions at the site of inoculation, saponin was considered more practical than the oil-based or bacterial adjuvants for the elicitation in cattle of antibodies to purified soluble antigens.
Enhancement of host susceptibility to lethal endotoxin shock by staphylococcal pyrogenic exotoxin type C
Authors: Schlievert P M
Published: January 2020

Staphylococcal pyrogenic exotoxin (PE) ty pe C enhanced the susceptibility of rabbits to lethal shock by endotoxin by as much as 50,000-fold. A graph of log PE type C dose used for pretreatment versus log 50% lethal dose of endotoxin gave a straight line with a slope of approximately -1. Rabbits that received PE type C alone showed fevers only, but those given both PE ty pe C and endotoxin showed initial fever followed by hypothermia, labored breathing, diarrhea, evidence of vascular collapse, and finally death. When a PE type C dose of 3 micrograms/kg was used, pretreatment of the animals with PE for 2 h before giving the endotoxin was required to obtain maximal susceptibility. However, when 15 micrograms of PE type C per kg was utilized, the endotoxin could be given before, concurrently, or after PE type C. The capacity of PE type C to prepare rabbits for enhanced susceptibility to endotoxin was lost after 24 to 48 h. Animals could be protected from enhanced susceptibility to endotoxin by prior immunization with either PE type C or endotoxin. However, 30% of the rabbits which were immunized with PE type C failed to develop immunity, and after three injections of PE type C, these animals developed gram-negative bacteremia and succumbed. In addition, rabbits with diarrhea initially, possibly caused by Pasteurella infection, died less than 24 h after a single injection of PE type C.
Myeloperoxidase-Mediated Oxidation of Methionine and Amino Acid Decarboxylation
Authors: Tsan Min-Fu
Published: January 2020

The myeloperoxidase (MPO)-mediated decarboxylation of amino acids and the MPO-mediated oxidation of methionine, two potential bactericidal mechanisms, were compared. In the presence of the MPO system (MPO, 50 mU/ml; H
2
O
2
, 0.1 mM; Cl
−
, 75 mM), 50% of alanine (0.1 mM) was decarboxylated, whereas only 5% of methionine (0.1 mM) was decarboxylated. In contrast, under similar conditions, 80% of methionine was oxidized to methionine sulfoxide. Once methionine was oxidized to methionine sulfoxide, it was decarboxylated (75%) by the MPO system. Methionine at 0.1 mM completely inhibited the decarboxylation of alanine, whereas alanine at a concentration 200 times that of methionine had no effect on the MPO-mediated oxidation of methionine. Sodium azide, an MPO inhibitor, inhibited the decarboxylation of alanine and the oxidation of methionine to the same extent. Tryptophan markedly inhibited the oxidation of methionine, whereas histidine stimulated it. Alanine, glycine, and taurine had no effect. In contrast, all of these amino acids and taurine markedly inhibited the MPO-mediated decarboxylation of alanine. NaN
3
, tryptophan, and methionine, which inhibited the MPO-mediated oxidation of methionine, also inhibited the killing of
Staphylococcus aureus
or
Klebsiella pneumoniae
by the MPO system; whereas histidine, alanine, and glycine, which did not inhibit the oxidation of methionine, had less or no effect on the killing of these two bacteria by the MPO system. Results suggest that methionine is preferentially oxidized to methionine sulfoxide by the MPO system. Once methionine is oxidized to methionine sulfoxide, it is then readily decarboxylated by the MPO system. The agent responsible for the oxidation of methionine may play an important role in the MPO-mediated killing of bacteria.
Effect of pyochelin on the virulence of Pseudomonas aeruginosa
Authors: Cox C D
Published: January 2020

A virulent isolate of Pseudomonas aeruginosa PAO1, which had been obtained from eight sequential intraperitoneal infections in mice compromised with iron and methotrexate, expressed greater lethality than the avirulent parent strain when both strains were injected into mice treated with iron. The present study demonstrates that pyochelin, a siderophore produced by P. aeruginosa, also increases the lethality of the virulent bacteria but not of the avirulent bacteria. Analysis of the growth and clearance of both virulent and avirulent strains in mice revealed that pyochelin increased the growth and lethality of virulent bacteria but only increased the survival of the avirulent bacteria. A streptomycin-dependent mutant of strain PAO1 (strd1) was used to demonstrate that pyochelin did not affect the clearance activity of mice. This strongly suggests that the effects of pyochelin in stimulating the persistence of avirulent bacteria and in increasing the lethality of virulent bacteria are due solely to the promotion of bacterial growth. Since the virulent bacteria were equivalent to the avirulent bacteria in utilizing pyochelin during in vitro growth in the presence of transferrin, it appears that the stimulation of growth by pyochelin allows the expression of additional virulence properties by the virulent bacteria.
Hemagglutination activity and colonization factor antigens I and II in enterotoxigenic and non-enterotoxigenic strains of Escherichia coli isolated from humans
Authors: Cravioto A , Scotland S M , Rowe B
Published: January 2020

We examined 205 enterotoxigenic strains of Escherichia coli for colonization factor antigens (CFA) I and II, using an immunodiffusion technique with specific antisera. A total of 36 strains of serogroups O63, O78, O114, O128, and O153 and 1 rough strain possessed CFA/I and gave a single precipitin line; 47 strains of serogroups O6, O8, O80, and O115 possessed CFA/II. The latter strains gave a major precipitin line (component 3) when tested with specific antisera prepared against strain E1392 or PB-176 (both E. coli O6.H16; biotype A). However, all 16 strains of E. coli O6.H16 belonging to biotype A gave a second precipitin line (component 1) when tested with both antisera. When CFA/II-positive strains were tested with a specific antiserum prepared against E. coli O6.H16 strains of biotype B or C, all strains gave component 3, but 16 of 17 strains of E. coli O6.H16 belonging to biotype B, C, or F gave a second precipitin line (component 2) not given by strains of biotype A. CFA/II-positive strains of serogroups other than O6 gave only component 3 in tests with all specific antisera. Nine enterotoxigenic strains of serotypes O7, O15, O25, O115, and O128 gave mannose-resistant hemagglutination of human or calf erythrocytes but lacked CFA/I or CFA/II. Although mannose-resistant hemagglutination was common in non-enterotoxigenic strains of E. coli, none of the non-enterotoxigenic strains possessed CFA/I or CFA/II; these strains included fecal strains of serogroups O6, O8, O63, and O78, fecal strains of enteropathogenic serogroups, and strains from extraintestinal sources.
Some characteristics of a urease-negative, temperature- sensitive strain of Bordetella bronchiseptica as a live, attenuated vaccine
Authors: Shimizu T , Ishikawa H
Published: January 2020

Strain ts-S34 of Bordetella bronchiseptica was treated jointly with two mutagens, nitrosoguanidine and UV irradiation, and a urease-negative (u-), temperature-sensitive (ts) mutant strain indistinguishable from Alcaligenes faecalis in its biochemical characteristics was isolated. However, the mutant isolated was a phase III organism. By repeating selection of smaller, hemolytic colonies observed among phase III colonies after prolonged incubation, a phase I organism, strain ts-S34.u-, was isolated. The u- and ts properties of the mutant strain were hereditarily stable, and heat-labile toxin production was very low. Growth in KCN broth and on Simmons citrate agar was reduced. Agglutinability of the strain against anti-S1 and anti-ts-S34.u- hyperimmune sera was as high (X20,480) as that of wild-type strain S1. The live ts-S34.u- strain vaccination protected guinea pigs from challenge exposure with 15,000 X the 50% lethal dose of virulent strain S1 of B. bronchiseptica. In these studies, it appeared that the ts-S34.u- strain has favorable properties, including a useful hereditary marker and low heat-labile toxin production, for use as a live attenuated vaccine.
Anatomical and immunological responses of rabbit gallbladders to bacterial infections
Authors: Gilman R H , Young C , Bulger R , Hornick R B , Greenberg B
Published: January 2020

To study the sequential morphological and immunological response of the rabbit gallbladder to bacterial infection and to compare the inflammatory responses with different pathogens, gallbladders were infected with Streptococcus faecalis and two strains of Escherichia coli, one of which produced enterotoxin. Gallbladder infection was produced either by intravenously injecting bacteria into rabbits with a small liver infarct or by injecting bacteria directly into the gallbladder of normal rabbits. The percentage of gallbladders infected intravenously with a nonenterotoxigenic E. coli strain was 86% at 1 week, 70% at 3 weeks, and 15% at 6 weeks. Epithelial necrosis and leukocyte infiltration were prominent 1 week after infection. At 3 and 6 weeks after infection, there was crypt distortion and increased mucus secretion in the epithelium as shown by periodic acid-Schiff staining. The lamina propria was infiltrated with mononuclear cells, many of which were plasma cells. Myofibroblasts (contractile fibroblasts) were also identified on transmission microscopy, In addition to these changes, toxigenic E. coli produced subepithelial capillary dilation in the villus core. Morphological changes (excluding toxin-associated changes) were related to the duration of infection rather than to the specific species of infecting bacteria. Infected gallbladders studied by immunofluorescence had a greater than 50-fold increase in plasma cells compared with control cells. In addition, the number increased with the duration of infection. Immunoglobulin A cells were the major cell type in gallbladders infected by intravesical injection, whereas immunoglobulin G cells predominated in gallbladders infected intravenously. The gallbladder appears to mount a local immune response to bacterial infection.
Bacteriophages in Sputum of Patients with Bronchopulmonary
Pseudomonas
Infections
Authors: Tejedor Carmen , Foulds John , Zasloff Michael
Published: January 2020

Pseudomonas aeruginosa
bacteriophages were recovered from the sputum of patients with chronic
P. aeruginosa
bronchopulmonary infection. Host ranges of some of the phages are presented.
Evaluation of a phenotypic revertant of the A/Alaska/77-ts-1A2 reassortant virus in hamsters and in seronegative adult volunteers: further evidence that the temperature-sensitive phenotype is responsible for attenuation of ts-1A2 reassortant viruses
Authors: Tolpin M D , Clements M L , Levine M M , Black R E , Saah A J , Anthony W C , Cisneros L , Chanock R M , Murphy B R
Published: January 2020

In a previous study, a seronegative child to whom attenuated A/Alaska/77-ts-1A2 virus was administered (37 degrees C shutoff temperature for plaque formation) shed virus with an altered temperature-sensitive (ts) phenotype (40 degrees C shutoff temperature) (Murphy et al., Ann. N.Y. Acad. Sci. 354:172-182, 1980; Tolpin et al., Virology 112:505-517, 1981). This ts+ virus (FV1319) was evaluated for its level of replication in hamsters and for its virulence for humans. In hamsters, FV1319 ts+ virus replicated to the same level in the nasal turbinates as that of which the A/Alaska/77 wild-type virus replicated, but its replication in the lungs was reduced 40-fold. In contrast, the A/Alaska/77-ts-1A2 reassortant achieved a titer in hamster nasal turbinates that was significantly lower (P less than 0.005) than those achieved by the wild-type and the FV1319 viruses; the A/Alaska/77-ts-1A2 reassortant was not recoverable from the lungs. In seronegative adult volunteers, the pattern of replication of the FV1319 virus was similar to that of the A/Alaska/77 wild-type virus. The illness induced by the FV1319 ts+ virus was also similar to that caused by the wild-type virus. In contrast, the A/Alaska/77-ts-1A2 reassortant was satisfactorily attenuated in adult volunteers. These results suggest that attenuation of the A/Alaska/77-ts-1A2 reassortant virus in humans is a function of the ts phenotype: loss of this phenotype restored virulence. The ability of the A/Alaska/77-ts-1A2 reassortant to lose its ts phenotype and regain virulence during growth in a permissive host limits the usefulness of the ts-1A2 reassortants as vaccine viruses for humans.
Production, purification, and characterization of the fimbrial adhesive antigen F41 isolated from calf enteropathogenic Escherichia coli strain B41M
Authors: de Graaf F K , Roorda I
Published: January 2020

Enterotoxigenic Escherichia coli strains of serogroups O9 and O101 produce an adhesive antigen, provisionally designated as F41. Production of the F41 antigen was shown to be dependent on the composition of the growth medium. A minimal salts medium or Minca medium was the most suitable medium to obtain a high production. The biosynthesis of the F41 antigen was repressed at 18 degrees C or in the presence of L-alanine. The F41 antigen was isolated from the bacteria by mechanical detachment, concentrated by precipitation with ammonium sulfate, and purified gel filtration on Sepharose CL-4B and treatment with deoxycholate. The purified F41 was composed of protein subunits with an apparent molecular weight of 29,500 on sodium dodecyl sulfate-polyacrylamide gels. The isoelectric point of the antigen was 4.6. The N-terminal amino acid sequence was determined. The F41 antigen had strong hemagglutinating activity with guinea pig and human group A erythrocytes and weaker hemagglutinating activity with horse and sheep erythrocytes. In immunoelectrophoresis at pH 8.4 the purified antigen migrated to the anode. Purified F41 antigen has a filamentous structure with an average diameter of 3.2 nm.
Immunoglobulin A and secretory immunoglobulin A antibodies to purified type 1 Klebsiella pneumoniae pili in human colostrum
Authors: Davis C P , Houston C W , Fader R C , Goldblum R M , Weaver E A , Goldman A S
Published: January 2020

Immunoglobulin A antibodies with binding specificity for purified Klebsiella pneumoniae type 1 pili were detected by an enzyme-linked immunosorbent assay in 20 of 21 human samples (95%). The concentrations of secretory immunoglobulin A antibody in colostrum directed against the pili were calculated by comparison of experimental enzyme-linked immunosorbent assay values with values obtained from known secretory immunoglobulin A concentrations. The presence of antibodies to K. pneumoniae type 1 pili was confirmed by double diffusion-gel studies with selected specimens of colostrum. This study shows that in the majority of human colostral samples examined, secretory immunoglobulin A antibodies with specificity for K. pneumoniae type 1 pili can be commonly found in variable, but frequently high, concentrations.
Alterations in nucleotide content of human lung fibroblasts infected with Mycoplasma pneumoniae
Authors: Upchurch S , Gabridge M G
Published: January 2020

The nucleotide content of normal MRC-5 human lung fibroblasts and fibroblasts infected with Mycoplasma pneumoniae PI 1428 was determined. Nucleotides from control and infected fibroblasts were extracted with 5% trichloracetic acid. After neutralization of the extracts, the nucleotides in the extracts were separated by anion-exchange chromatography. Significant differences were found between the nucleotide content of the control and infected cells. Nucleotide triphosphate levels were twofold higher in the control fibroblasts than in the infected fibroblasts 4 h after the initiation of infection. At the same time, nucleotide diphosphate and monophosphate levels were higher in the infected fibroblasts than in the control fibroblasts. Determination of the energy charge ratio for each set of nucleotides (adenosine, guanosine, cytidine, and uridine) demonstrated a shift of nucleotide content in the infected fibroblasts. Immediately after infection, the energy charge for each set of nucleotides was higher for the control fibroblasts than it was for the infected fibroblasts. This pattern continued throughout the infection period with only minor exceptions. The work presented here indicates a loss of energy charge in fibroblasts infected with M. pneumoniae and may help to explain some of the metabolic changes and cell damage which accompany infection.
Cloning of genes determining the production of mannose-resistant fimbriae in a uropathogenic strain of Escherichia coli belonging to serogroup O6
Authors: Clegg S
Published: January 2020

Chromosomal DNA from a uropathogenic strain of Escherichia coli was partially digested with the restriction enzyme EcoRI. The partial digests were ligated into a cosmid containing an ampicillin-resistant determinant and packaged into lambda phage particles. An ampicillin-resistant transductant of E. coli HB101 was found to possess mannose-resistant hemagglutinating activity associated with a 50-kilobase-pair plasmid. Subcloning of the mannose-resistant fimbrial genes revealed that the genetic determinants were encoded by a 6.9-kilobase-pair DNA fragment of a recombinant plasmid. Chimeric plasmids smaller in size were unable to transform E. coli to fimbrial production. Physical maps of the recombinant plasmids were prepared showing restriction endonuclease sites within the inserted DNA fragments. The hemagglutinating activities of the wild-type strain and of the recombinant derivative were compared. Both strains agglutinated human erythrocytes in the presence of D-mannose to the same degree and also failed to produce fimbriae after incubation at 18 degrees C. Also, both strains were agglutinated by antifimbrial serum at a high titer, whereas no such activity was observed when a strain of E. coli which did not possess a plasmid was used.
Surface receptors for serum albumin in group C and G streptococci show three different types of albumin specificity
Authors: Wideback K , Kronvall G
Published: January 2020

A total of 100 bacterial strains were tested for binding uptake of radiolabeled albumin preparations from 15 mammalian species. Three types of surface structures with specific binding sites for albumin were defined. A previously described receptor for albumin was separated into type a in Streptococcus equisimilis strains and in human group G streptococcal strains and type b in bovine group C streptococci. A new type of albumin receptor, type c, was found in Streptococcus dysgalactiae strains, the only receptor type so far with high affinity for bovine serum albumin. Type of albumin receptor correlated with bacterial species. The three receptor types showed high binding capacities; 2 X 10(8) bacterial organisms bound from 5 to 16 micrograms of albumin. All types of albumin receptors were stable to heat treatment at 80 degrees C for 5 min, but susceptible to both pepsin and trypsin treatment. Bacteria-bound albumin preparations were eluted at various concentrations of KSCN, reflecting differences in affinity. Up to 500 micrograms of human fibrinogen or polyclonal human immunoglobulin G had no inhibitory effect on the uptake of albumin, indicating a separate molecular localization of receptors for these proteins.
Mitogenic activity of Sindbis virus and its isolated glycoproteins
Authors: Goodman-Snitkoff G , McSharry J J
Published: January 2020

Purified preparations of Sindbis virus, a member of the togavirus family, are mitogenic for lymphocytes from a number of different mouse strains. Cell separation techniques, as well as studies using lymphocytes from the congenitally athymic BALB/c nu/nu mouse, showed that Sindbis virus is a T-cell-independent B-cell mitogen. Additionally, the envelope glycoproteins of Sindbis virus, isolated by Triton X-100 extraction and butanol precipitation, stimulated lymphocytes to incorporate five times as much [3H]thymidine into their DNA as did the Sindbis virion. These results are similar to those previously reported for vesicular stomatitis virus and herpes simplex virus types I and II and for the purified glycoproteins of vesicular stomatitis virus and influenza virus.
Characterization of the lipopolysaccharide from Vibrio cholerae 395 (Ogawa)
Authors: Kabir S
Published: January 2020

The chemical structure and biological properties of the lipopolysaccharide (LPS) from Vibrio cholerae 395 (Ogawa), isolated by the phenol-water procedure, were studied. Upon acid hydrolysis, the LPS was split into its polysaccharide and lipid A moieties. The polysaccharide contained both neutral (glucose, heptose, fructose) and amino (glucosamine, quinovosamine) sugars. The LPS contained the acid-labile amino sugar, 4-amino-arabinose, which was absent in the Inaba serotype of V. cholerae. The LPS differed from the LPSs of Enterobacteriaceae by the absence of 2-keto-3-deoxyoctonate and the presence of fructose. Analysis of the methylated polysaccharide by gas-liquid chromatography and mass spectrometry showed that it had a branched structure with glucose and heptose residues primarily appearing at the nonreducing-end groups. Interactions with lectins, concanavalin A. and wheat germ agglutinin suggested that terminal glucose residues were alpha linked, whereas terminal glucosamine residues were connected by alpha-1,3 linkages. The major fatty acids of the LPS were C14:0, C16:0, C12h:0, and C14h:0 compounds, of which only the C14h:0 were amide linked, the remainder being ester linked to the backbone. Biological studies showed that the LPS possessed endotoxic properties such as lethality, pyrogenicity, limulus lysate gelation, and ability to induce non-specific resistance to infection. Thus, the LPS from V. cholerae 395 (Ogawa) possessed both common and distinct features as compared with the LPSs from the Enterobacteriaceae.
Nocardia brasiliensis: mycetoma induction and growth cycle
Authors: Conde C , Melendro E I , Fresán M , Ortiz-Ortiz L
Published: January 2020

The capacity of Nocardia brasiliensis to induce mycetoma in BALB/c mice in the absence of adjuvants was studied. Whether the virulence of N. brasiliensis varied in the different phases of its growth cycle was also determined. The results showed that N. brasiliensis suspended in 0.15 M NaCl and injected into the footpads of mice were able to induce mycetoma after only 14 days of infection, as evidenced by histological studies. Data are also presented indicating that the virulence of N. brasiliensis did not vary during the different phases of its growth curve. The differences in virulence reported between N. brasiliensis and other nocardiae are discussed and explained in terms of several variables in the experimental designs used: among these variables are mouse genotype, route of inoculation, and model for determining virulence.
Effect of purified protein derivative and sonicates of Mycobacterium leprae and Mycobacterium bovis BCG on thromboplastin response in human monocytes in vitro
Authors: Lyberg T , Closs O , Prydz H
Published: January 2020

Human monocytes isolated from peripheral blood responded with increased thromboplastin expression upon stimulation in vitro with three mycobacterial antigens: tuberculin purified protein derivative and sonicates of Mycobacterium boviS BCG and Mycobacterium leprae. The stimulating principle of mycobacteria is probably a cell wall constituent since crude extracts of cell walls were 2.5 to 25 times more potent in stimulating thromboplastin synthesis than were whole sonicates. This thromboplastin response was inhibited by inhibitors of RNA and protein synthesis, dexamethasone, and agents that caused elevation of intracellular cyclic AMP. The presence of lymphocytes did not enhance the monocyte thromboplastin response significantly during the first 24 h of incubation. For M. bovis BCG and M. leprae sonicates, the thromboplastin response correlated with general activating effects measured by determining the release of lysozyme and beta-glucuronidase. The role of thromboplastin in chronic inflammatory reactions is discussed.
Comparison of the calcium ionophore and phorbol myristate acetate on the initiation of the respiratory burst in human neutrophils
Authors: DeChatelet L R , Lees C J , Walsh C E , Long G D , Shirley P S
Published: January 2020

We investigated the ability of the calcium ionophore A23187 and phorbol myristate acetate to elicit a respiratory burst in human neutrophils. In general, the ionophore was considerably more potent than phorbol myristate acetate in generating a chemiluminescent response and slightly less active in generating superoxide anion. In contrast, the ionophore caused a much smaller stimulation of glucose oxidation via the hexose monophosphate shunt. This relative inability of the ionophore to stimulate the shunt could not be ascribed to an effect on glucose transport or to a direct inhibition of any of the enzymes involved in glucose metabolism. These data suggest that different stimuli may have markedly different effects on various activities associated with the respiratory burst and emphasize the necessity for measurement of more than one parameter to assess the oxidative metabolism of the neutrophil.
Properties of synthetically produced Escherichia coli heat-stable enterotoxin
Authors: Klipstein F A , Engert R F , Houghten R A
Published: January 2020

The properties of a synthetically produced peptide composed of the same primary structure of 18 amino acids described for human Escherichia coli heat-stable enterotoxin were compared with those of purified heat-stable toxin obtained by bacterial growth. The dosage required to evoke fluid secretion in the suckling mouse and rat ligated ileal loop assays was the same for both toxins. The antigenicity of the two toxins was similar when assayed by enzyme-linked immunosorbent assay with hyperimmune antiserum to either toxin. The secretory effect of the two toxins in the suckling mouse assay was seroneutralized by the same dilutions of hyperimmune antiserum to either toxin. Immunization of rats with the synthetic toxin coupled to a large-molecular-weight carrier raised serum and mucosal antitoxin responses which provided protection against challenge with either the synthetic or biological toxin as well as with viable heat-stable enterotoxin-in-producing organisms. These observations indicate that synthetically produced heat-stable toxin has the same properties as the toxin derived by bacterial culture. The availability of the more readily made synthetic form of heat-stable toxin should facilitate the production of a vaccine based on cross-linking this toxin with either the heat-labile toxin or its nontoxic B subunit.
Maturation In Vivo of
Schistosoma mansoni
Schistosomula After Culture In Vitro with Granulocytes and Antibody
Authors: Dessein Alain , Butterworth Anthony E. , Vadas Mathew A. , David John R.
Published: January 2020

Seven experiments were carried out to test the relationship between the morphological assay for damage to schistosomula in vitro with toluidine blue and the loss of the ability of damaged organisms to mature in vivo. Schistosomula were prepared by penetration of rat skin and cultured for 12 to 38 h in the presence of various combinations of purified human eosinophils or neutrophils and heat-inactivated human antischistosomular serum. Samples were scored for microscopically detectable damage, and the remaining organisms were injected intravenously into normal mice. These mice were perfused after 5.5 to 7 weeks, and the recovery of adult worms was determined. After culture of schistosomula in medium alone, between 8.4 and 32.7% of injected organisms matured into adult worms. There was no significant difference in the capacity of freshly prepared and cultured schistosomula to mature in vivo. Schistosomula cultured with antibody alone showed no significant damage in vitro, and in only one of seven experiments was there a significant (35%) reduction compared with the medium controls in their capacity to mature in vivo. Schistosomula cultured with neutrophils alone or eosinophils alone showed no significant damage in vitro and no loss of viability in vivo. Schistosomula cultured with neutrophils and antibody showed a 28% reduction in recovery in one experiment but an increase in recovery (12 and 46%) in two other experiments. In contrast, schistosomula cultured with eosinophils and antibody showed evidence of both marked damage in vitro (22 to 93% dead organisms) and loss of viability in vivo (26 to 98% reduction in recovery) in all seven experiments. These findings justify the use of the toluidine blue morphological assay as an estimate of irreversible damage to schistosomula and confirm that human eosinophils and neutrophils differ markedly in their capacity to mediate antibody-dependent damage in vitro.
Gamma-irradiated scrub typhus immunogens: development of cell-mediated immunity after vaccination of inbred mice
Authors: Jerrells T R , Palmer B A , Osterman J V
Published: January 2020

Mice immunized with three injections of gamma-irradiated Karp strain of Rickettsia tsutsugamushi were evaluated for the presence of cell-mediated immunity by using delayed-type hypersensitivity, antigen-induced lymphocyte proliferation, and antigen-induced lymphokine production. These animals also were evaluated for levels of circulating antibody after immunization as well as for the presence of rickettsemia after intraperitoneal challenge with viable Karp rickettsiae. After immunization with irradiated Karp rickettsiae, a demonstrable cell-mediated immunity was present as evidenced by delayed-type hypersensitivity responsiveness, lymphocyte proliferation, and production of migration inhibition factor and interferon by immune spleen lymphocytes. Also, a reduction in circulating rickettsiae was seen in mice immunized with irradiated rickettsiae after challenge with 1,000 50% mouse lethal doses of viable, homologous rickettsiae. All responses except antibody titer and reduction of rickettsemia were similar to the responses noted in mice immunized with viable organisms. Antibody levels were lower in mice immunized with irradiated rickettsiae than in mice immunized with viable rickettsiae. Furthermore, mice that were immunized with viable rickettsiae demonstrated markedly lower levels of rickettsemia after intraperitoneal challenge compared with either mice immunized with irradiated rickettsiae or nonimmunized mice.
Shigella dysenteriae 1 cytotoxin: periplasmic protein releasable by polymyxin B and osmotic shock
Authors: Donohue-Rolfe A , Keusch G T
Published: January 2020

Treatment of Shigella dysenteriae 1 either with the antibiotic polymyxin B or by osmotic shock resulted in the release of 80 to 90% of the cytotoxin activity of the organism. Under the conditions employed, the release of toxin activity was accompanied by the appearance of a periplasmic enzyme, 5'-nucleotidase. There was no significant release of cytoplasmic contents, assessed by measurement of glucose-6-phosphate dehydrogenase activity. The release of cytotoxin and 5'-nucleotidase by polymyxin B were both dependent on the duration of incubation with, and the concentration of, the antibiotic. In terms of specific activity (cytotoxin activity per milligram of protein), the polymyxin B and osmotic shock extracts were 20- to 30-fold more active than crude toxin preparation derived from a whole-cell lysate. The data strongly support a periplasmic location for Shiga cytotoxin and the utility of the polymyxin B extraction to obtain starting material for toxin purification.
Evidence of a soluble substrate for the receptor-destroying enzyme of influenza C virus
Authors: O'Callaghan R J , Labat D D
Published: January 2020

Influenza C virus contains a hemagglutinin and a receptor-destroying enzyme (RDE) whose specificities remain undetermined. In rat serum, there is a molecule that binds specifically to C virus, inhibiting its hemagglutinin. The complex between C virus and the rat serum inhibitor (RSI) was determined to be stable at 4 degrees C, but was disrupted within 20 to 90 min at 23 or 37 degrees C. Virus emerged from the complex with numerous functions intact, whereas the RSI at this point was inactivated, i.e., incapable of further inhibitory reactions with C virus. RSI could not be inactivated at these temperatures by nonviral components of allantoic fluid of infected chicken embryos; however, RSI inactivation was achieved by preparations of sucrose gradient-purified virus. Neutralization of viral hemagglutination activity with antiviral antibody protected the RSI from inactivation. RSI inactivation occurred at temperatures at which the viral RDE was active, and inhibition of viral RDE by periodate treatment sharply reduced the ability of virus to inactivate the RSI. One interpretation of the data suggests that RSI is a receptor analog reactive with both the hemagglutinin and RDE of C virus and that RSI inactivation is an assay of influenza C viral RDE.
Serum Resistance of
Yersinia enterocolitica
Expressed in Absence of Other Virulence Markers
Authors: CHIESA Claudio , Bottone Edward J.
Published: January 2020

Serum resistance of
Yersinia enterocolitica
after growth at 37 and 25°C appears to be specific for serogroup O:3 and appears to be expressed even in the absence of other phenotypic virulence-associated markers, such as the presence of V antigen, autoagglutination, and calcium dependency after growth at 37°C.
Turnover in the transferrin iron pool during the hypoferremic phase of experimental Neisseria meningitidis infection in mice
Authors: Letendre E D , Holbein B E
Published: January 2020

Mouse transferrin was used to specifically label the plasma transferrin iron pool for studies of iron kinetics in normal mice and infected mice during the hypoferremic phase of experimental meningococcal infection. The plasma transferrin iron pool of normal mice was found to be very dynamic, with a half-life of iron in the pool of 0.7 h. Iron left the plasma pool, entered the bone marrow, and was released into the blood in erythrocytes. Iron from the transferrin pool also entered the liver and spleen and was presumably in the reticuloendothelial system components of these organs. Most of the iron that had been supplied as transferrin iron was found in erythrocytes by 48 h after injection. Studies with mice infected with Neisseria meningitidis strain M1011 revealed similar kinetics for transferrin iron. There was no redistribution of iron within the various iron pools as a result of infection. Iron turnover in the plasma transferrin pool during the hypoferremic phase was similar to control rates, and iron leaving the pool entered its normal erythroid compartments. The lack of accelerated turnover of plasma iron and the finding that plasma iron was not rerouted to storage compartments during the hypoferremic phase provided good evidence that lactoferrin and leukocytic endogenous mediator were not directly involved in redirecting transferrin iron. Our evidence has implicated an impaired return of reticuloendothelial system-processed iron to the transferrin pool during the hypoferremic response. This appears to be a logical point in the erythroid iron cycle for host-mediated iron sequestration, as the reticuloendothelial system is involved in iron storage and may regulate iron levels in the plasma transferrin pool under normal conditions.
Phagocytosis and Killing of
Francisella tularensis
by Human Polymorphonuclear Leukocytes
Authors: Löfgren Sture , Tärnvik Arne , Bloom Gunnar D. , Sjöberg Wigert
Published: January 2020

Bacteria of a wild strain of
Francisella tularensis
were less efficiently killed by human polymorphonuclear leukocytes than were bacteria of an attenuated strain. This finding was explained to some extent by a less efficient phagocytosis, but bacteria of the wild strain also seemed to be more resistant to killing after ingestion.
Enhancement of Natural Resistance to Influenza Virus in Lipopolysaccharide-Responsive and -Nonresponsive Mice by
Propionibacterium acnes
Authors: Gangemi J. David , Hightower James A. , Jackson Ronnie A. , Maher Michael H. , Welsh Marcia G. , Sigel M. Michael
Published: January 2020

Lipopolysaccharide-responsive C3H/HeN mice were rendered resistant to a mouse-adapted strain of influenza (Aichi, H
3
N
2
) virus when
Propionibacterium acnes
was given either intranasally or intraperitoneally several days before virus infection. The time of
P. acnes
treatment was important since no protection was demonstrated when this agent was given either on the same day as or several days after virus challenge. In contrast, lipopolysaccharide-nonresponsive C3H/HeJ mice were not protected when
P. acnes
was administered intranasally at any time before infection; however, protection was demonstrated when
P. acnes
was given by the intraperitoneal route. Depending on the route of inoculation,
P. acnes
induced several distinctive immunological responses in the lungs of both C3H/HeN and C3H/HeJ mice. Intranasal inoculation was more effective in activating pulmonary macrophages in C3H/HeN than in C3H/HeJ mice. In contrast, intraperitoneal inoculation activated pulmonary natural killer cells in both mouse lines but did not activate pulmonary macrophages.