bluuebunny/crossref_metadata_2025 huggingface dataset:
Vi Antigen from
Salmonella typhosa
and Immunity Against Typhoid Fever I. Isolation and Immunologic Properties in Animals
Authors: Wong K. H. , Feeley John C. , Northrup R. S. , Forlines M. E.
Published: January 2020

The role of Vi antigen in human immunity against typhoid fever has been debated for decades. Circumstantial evidence indicates that Vi antigen may play a role in pathogenicity and immunity. A Vi preparation was isolated from
Salmonella typhosa
, the causative organism in human typhoid fever, by a mild precipitation method. It was significantly more potent in animal studies than preparations from
Citrobacter
used in the past for human study and less toxic than conventional typhoid vaccines. With this antigen, the role of Vi antigen in human protection is now being investigated.
Relationship of Antimicrobial Cellular Immunity to Delayed Hypersensitivity in Listeriosis
Authors: Osebold John W. , Pearson Leonard D. , Medin Nicolas I.
Published: January 2020

The relationship of antimicrobial cellular immunity to delayed hypersensitivity (DH) was studied in mice antigenically stimulated by living
Listeria monocytogenes
confined to diffusion chambers in peritoneal cavities or by subcutaneous inoculation of sublethal doses of the organism. Mice showed DH reactions when tested 6 days after inoculation, and reactions were positive for at least 90 days in some mice. DH also became established when the mice were stimulated by antigens diffusing from peritoneal chambers containing viable
Listeria
. Mice were categorized as DH positive or DH negative if they developed more or less than a 5% increase in foot volume 24 h after the injection of
Listeria
antigen. Some antigenically stimulated mice did not elicit the DH reaction. Consequently, the animals were arranged as immunized groups (DH positive and DH negative) and
Listeria
chamber implant groups (DH positive and DH negative). When challenged with
L. monocytogenes
, all four groups were significantly resistant as compared with controls. Thus, the in vivo tests for immunity and DH did not show direct correlation. The results suggested that antimicrobial cellular immunity can occur as a phenomenon independent of DH. Evidence for antimicrobial cellular immunity as the principle mechanism of resistance in murine listeriosis is discussed with consideration for possible heterogeneity of function by thymus-derived lymphocytes.
Stability of the Rabbit Immunogenic Marker of RA 27/3 Rubella Vaccine Virus After Human Passage
Authors: Linnemann Calvin C. , Hutchinson Leslie , Rotte Thomas C. , Hegg Marion E. , Schiff Gilbert M.
Published: January 2020

Rabbits were inoculated intravenously with “wild” rubella virus, RA 27/3 rubella vaccine virus, or rubella virus isolated from recipients of RA 27/3 vaccine. Rabbits receiving “wild” virus developed rubella hemagglutination inhibition antibody, and those receiving vaccine virus did not. One of the five reisolates tested produced a low transient antibody response in two of the five rabbits inoculated with this strain. The study indicates that the rabbit immunogenic marker after intravenous injection can be used to determine if a rubella virus isolated from a patient is of “wild” or vaccine origin. There was no significant change in the reduced immunogenicity characteristics of the RA 27/3 vaccine virus after human passage.
Immune-Mediated Emigration of Neutrophils into the Lumen of the Small Intestine
Authors: Bellamy James E. C. , Nielsen N. Ole
Published: January 2020

By using pigs sensitized to bovine serum albumin (BSA), it was found that exposure of the intestinal mucosa to BSA induced, in 4 h, the emigration of large numbers of neutrophils into the intestinal lumen. This response was specific for the immunizing antigen and could be transferred to nonimmune animals with immune serum. The emigration of neutrophils through the intestinal mucosa was not accompanied by the edema, hemorrhage, and thrombosis which were apparent after intracutaneous inoculation of BSA into the same animals. Twenty-four hours after a 4-h mucosal exposure to BSA, the intestinal mucosa showed no evidence of neutrophil emigration nor any other abnormal features. These observations suggested that emigration of neutrophils into the intestinal lumen can be a specific, antibody-mediated immune response which occurs in the absence of intestinal injury. Possible relationships between immune-mediated enteroluminal emigration of neutrophils, neutrophil production, and a protective role for the neutrophil in the intestinal lumen were considered.
Enhancement Activity of Homologous Anti-Staphylococcal Sera in Experimental Staphylococcal Synovitis of Chicks: A Possible Role of Immune Adherence Antibodies
Authors: Forget A. , Meunier L. , Borduas A. G.
Published: January 2020

Homologous anti-staphylococcal sera passively potentiated the development of experimental staphylococcal synovitis infection of chickens. These antisera obtained from chickens hyperimmunized with live
Staphylococcus aureus
were selected according to their immune adherence and agglutinating properties. While the agglutinins rose steadily for 11 weeks during the hyperimmunization schedule, the immune adherence antibody titers reached a peak at 8 weeks and dropped to almost zero 2 to 4 weeks later. The enhancement activity of these antisera was associated with a relatively high level of immune adherence antibodies and seemed not to be correlated with their agglutinin titers.
Control of Interferon, Migration Inhibition Factor, and Expression of Virus Capsid Antigens in Burkitt's Lymphoma-Derived Cell Lines: Role of
l
-Arginine
Authors: Archer D. L. , Young B. G.
Published: January 2020

The effects of an arginine-utilizing mycoplasma,
Mycoplasma arginini
, and of varying levels of arginine in the growth media of the Epstein-Barr virus (EBV)-containing EB1 and EB3 cell lines, and the EBV-free RPMI 1788 cell line, were studied.
l
-Arginine, at a concentration of 0.1 mM in the growth medium, led to a reduction of the EBV capsid antigen content of the EB1 and EB3 cells lines as determined by indirect immunofluorescence, and
M. arginini
infection enhanced this reduction. The synthesis of two immune products, interferon and macrophage migration inhibition factor, was enhanced by growing the cell lines in medium containing arginine at a concentration of 0.1 mM, but the RPMI 1788 cell line produced much less of both products than EB1 or EB3 under these conditions. Infection of the cell lines with
M. arginini
reduced the amount of interferon produced and completely inhibited macrophage migration inhibition factor synthesis. The addition of arginine to a final concentration of 0.6 mM in the growth medium caused a dual effect: the EB1 and EB3 cell lines maintained the original level of EBV capsid antigens, even when infected with
M. arginini;
immune product synthesis was greatly reduced or completely inhibited by the addition of arginine, and
M. arginini
infection caused no further reduction of immune product synthesis.
Enhanced In Vitro Phagocytosis of
Listeria monocytogenes
by Human Monocytes in the Presence of Ampicillin, Tetracycline, and Chloramphenicol
Authors: Adam Dieter , Schaffert Wolfgang , Marget Walter
Published: January 2020

An in vitro system was developed to test the phagocytic activity of human macrophages grown from blood monocytes in the presence of the antibiotics ampicillin, tetracycline, and chloramphenicol.
3
H-labeled
Listeria monocytogenes
served as test organism. Subinhibitory amounts of the antibiotics enhanced the phagocytic activity significantly (
P
< 0.025). Macrophages pretreated with the drugs in identical concentrations showed the same phagocytic activity as control cells in the absence of the drugs. Because the drug concentrations used were similar to those that may be attained in man at certain places of inflammation, enhanced phagocytosis in the presence of antibiotics may have clinical significance.
Serological Evidence of Infection by Pichinde Virus Among Laboratory Workers
Authors: Buchmeier Michael , Adam Ervin , Rawls William E.
Published: January 2020

A total of 44 laboratory workers were examined for serological evidence of infection by Pichinde virus. Complement-fixing antibodies were detected in the sera of 6 of 13 (46%) persons working with the virus but not among 31 persons not working with the virus. Serological evidence of infection was greatest among those workers exposed to concentrated virus. No distinct illness could be associated with the infection.
Physicochemical Fractionation of Extracellular Cornea-Damaging Proteases of
Pseudomonas aeruginosa
Authors: Kreger Arnold S. , Griffin Olwen K.
Published: January 2020

Fractionation of the culture supernatant fluids of a cornea-virulent strain of
Pseudomonas aeruginosa
by ammonium sulfate precipitation, diafiltration, isoelectric focusing, ion-exchange chromatography, gel filtration, and sucrose density gradient centrifugation failed to separate the rabbit cornea-damaging activity and the in vitro protease activity of the preparations. Three proteases having similar molecular weights (approximately 20,000) and isoelectric points of approximately 4.6, 5.8, and 8.8 were obtained free of detectable amounts of other known extracellular pseudomonal enzymes. Heating a mixture of the three proteases for 15 min at 80 C resulted in complete loss of protease and cornea-damaging activities. The sterile culture filtrate of a nonproteolytic but lethal toxin-producing strain of
P. aeruginosa
did not contain cornea-damaging activity. Cultivation of the proteolytic strain in broth containing 4.7% ammonium sulfate yielded a culture supernatant fluid free of protease and cornea-damaging activities. The results obtained support the conclusion that a cornea-virulent strain of
P. aeruginosa
can produce, in vitro, at least three different extracellular proteases capable of eliciting rapid and extensive damage to rabbit corneas.
Studies on the Effect of Starvation on Mycobacteria
Authors: Nyka W.
Published: January 2020

Ten cultures of
Mycobacterium tuberculosis
, one of
Mycobacterium kansasii
(nonsignificant), and one of
Mycobacterium phlei
were submitted to starvation. As a result they lost first their acid fastness and then all other staining affinities but, in this chromophobic state, they survived for at least 2 years and, after that time, produced cultures of acid-fast bacilli when transferred onto nutrient media. Chromophobic tubercle bacilli similar to those produced experimentally had previously been demonstrated in caseous lesions of lungs surgically removed from patients under chemotherapy. Since it has been shown that experimentally produced chromophobic bacilli can recover their original biological properties, the opinion is warranted that, under suitable conditions, those in the lung could also become reactivated and cause a relapse of the disease.
Cross-Protection Between Group B Arboviruses: Resistance in Mice to Japanese B Encephalitis and St. Louis Encephalitis Viruses Induced by Dengue Virus Immunization
Authors: Tarr G. Crissman , Hammon William McD.
Published: January 2020

Albino Swiss mice, immunized with any of several types and strains of dengue viruses, were afforded substantial protection against peripheral Japanese B encephalitis or St. Louis encephalitis virus challenge. Dengue-2 (New Guinea “C”)-immunized mice showed, 10 and 20 weeks after immunization, undiminished resistance with concomitant cyclophosphamide treatment and virus challenge. Examination of the effects of immunization on Japanese B encephalitis virus pathogenesis, after virus challenge with concomitant cyclophosphamide treatment, indicated that protection was associated with decreased viremia and virtually no virus replication in the brain as compared with controls. These effects could be demonstrated before detection of any neutralizing antibody to the challenge virus. From the applied aspect, the data support the hypothesis, based on epidemiological evidence and experiments in hamsters, that prior exposure of man to dengue viruses can confer some degree of protection against Japanese B encephalitis or St. Louis encephalitis disease.
Action of
Escherichia coli
Enterotoxin: Adenylate Cyclase Behavior of Intestinal Epithelial Cells in Culture
Authors: Kantor Harvey S. , Tao Pearl , Wisdom Charlene
Published: January 2020

Heat-labile enterotoxin preparations obtained from two enteropathogenic strains of
Escherichia coli
of porcine and human origin were shown to stimulate adenylate cyclase activity of human embryonic intestinal epithelial cells in culture. Comparable results were also obtained when cholera toxin was used. The degree of enzyme stimulation was proportional to the concentration of enterotoxin. Similar preparations from two strains of non-enterotoxigenic
E. coli
had no effect on adenylate cyclase activity. Cells exposed to enterotoxin could be washed after 1 min of contact time without altering the subsequent course of maximum adenylate cyclase activity, which was maintained for at least 18 h at 37 C. During long periods (18 h) of tissue culture incubation, the determination of adenylate cyclase activity was 200- to 300-fold more sensitive than quantitating fluid accumulation in the adult rabbit ileal loop model. Decreasing the incubation time appreciably reduced the sensitivity of the epithelial cells to enterotoxin.
E. coli
enterotoxin is an effective activator of nonintestinal adenylate cyclase systems. Treatment of KB and HEp-2 cell lines with enterotoxin also resulted in significant enzyme stimulation. The intestinal epithelial cell tissue culture model provides a sensitive homogenous biological system for studying the response of intestinal adenylate cyclase to enterotoxin while eliminating the numerous cellular and tissue components present in the ligated ileal loop model.
Reduced Cell Yields of Mouse Cell Line Cultures After Exposure to Homogenates of Multiple Sclerosis Tissues
Authors: Carp Richard I. , Merz George S. , Licursi Pamela C.
Published: January 2020

The total cell yields in cultures of a mouse cell line termed “PAM” were reduced markedly after exposure to multiple sclerosis (MS) homogenates. The reductions were noted as early as the second subcultivation. The effect was produced by eight of eight MS brain homogenates and three of three MS spleen homogenates; the samples were from 10 MS cases. The reduction in total cell yields (compared to medium-treated cultures) was not observed after exposure to homogenates, six brain and two spleen, from eight non-MS cases. The effect was produced by the 50-nm filtrate of a pool of MS brain homogenates. Five of the MS-inoculated cultures were maintained for 18 subcultivations, and the reduction in total cell yields persisted throughout the series. The capacity to induce the reduction in cell yields was present in cell-free lysates from five of five MS cultures at passage 18, as was the capacity to cause a decrease in the polymorphonuclear neutrophils in the peripheral blood of mice. Calculation of the dilution effect (at least 10
18
-fold) that had occurred by the end of the passage series proved that the factor(s) causing the cell change in culture and the polymorphonuclear neutrophil change in mice had replicated in PAM cells.
Immune Response to Acute Otitis Media in Children I. Serotypes Isolated and Serum and Middle Ear Fluid Antibody in Pneumococcal Otitis Media
Authors: Sloyer John L. , Howie Virgil M. , Ploussard John H. , Amman Arthur J. , Austrian Robert , Johnston Richard B.
Published: January 2020

Seventy percent of pneumococci isolated from the middle-ear cavity of infants and children with acute otitis media were of one of the seven serotypes 1, 3, 6, 14, 18, 19, or 23. The immunological response in the serum and middle-ear fluid from otitis media caused by one of these serotypes was studied in 61 children by using either indirect hemagglutination or indirect fluorescent antibody tests, or both. Twenty-six of the patients had pneumococcal antibody present in the acute serum and 28 had it in the convalescent serum by at least one method. Thirteen of the 49 middle-ear fluids examined had antibody by the indirect fluorescent antibody technique. Serum pneumococcal antibody was found to reside predominantly in the immunoglobulin G or immunoglobulin M classes, whereas pneumococcal antibody with middle-ear fluid was found to be distributed equally among all three classes. Approximately 25% of the patients (16 of 61) had a positive immune response to their infection as evidenced by increased levels of pneumococcal antibody in the convalescent serum. The percentage of patients responding immunologically increased with age: 12% of infants less than 12 months showed a significant response, whereas 48% of children over 24 months responded.
New Streptococcal Serotypes Causing Pyoderma and Acute Glomerulonephritis Types 59, 60, and 61
Authors: Dillon Hugh C. , Dillon Mary S. A.
Published: January 2020

Three new streptococcal M serotypes, types 59, 60, and 61; have been described. They were first isolated from patients with pyoderma and acute glomerulonephritis (AGN), seen during epidemiological studies in Alabama. A possible antigenic relationship between types 59 and 61 was suggested by their T-agglutination reactions; a more specific T antiserum prepared for type 59 was useful in separating these two types, as well as other strains known to share T antigens 11, 12, and 5/27/44. On the basis of precipitin tests, a common antigenic determinant among types 59, 61, and 49 was suggested. This is of interest in view of the relation between these types and AGN. Type 59 has been relatively more widespread in distribution than type 61, but neither have been related to epidemic AGN. Type 60, first identified as “T-4,” thus being related to previously described M types which share this antigen, is of great interest in terms of the epidemiology of AGN. Most strains in our collection were recovered from patients with pyoderma and AGN or from their infected siblings. Recently, the type was found to be prevalent among patients with pyoderma and AGN seen in Trinidad. Data reported in relation to these new types further illustrate the dichotomy in M serotypes common to pyoderma and AGN on the one hand and those types found in collections of patients with pharyngitis on the other hand. The high rate of non-M-typable streptococci among pyoderma collections, encountered earlier, is best explained by lack of suitable reference antisera available for prevalent pyoderma serotypes.
Effects of Local Immunization with
Streptococcus mutans
on Induction of Salivary Immunoglobulin A Antibody and Experimental Dental Caries in Rats
Authors: Taubman Martin A. , Smith Daniel J.
Published: January 2020

The effect of local immunization with
Streptococcus mutans
on dental caries in conventional and gnotobiotic rats was studied. Injection of these animals with
S. mutans
strain 6715 incorporated into complete Freund adjuvant consistently resulted in the presence of antibody in saliva directed to this organism. This antibody was primarily of the immunoglobulin A class as demonstrated by specific antiglobulin augmentation and gel filtration of antibody activity. Serum antibody was also present. Five experiments have been completed in conventional rats and two in gnotobiotic animals. The immunized group of animals always had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. The numbers of lesions were also always lower in the immunized animals, suggesting a possible interference with the formation of new lesions in immunized animals. The reductions in dental caries and lesions were greater on smooth surfaces than on occlusal surfaces. which might be explained as interference with adherence phenomena demonstrated by
S. mutans.
It is proposed that salivary immunoglobulin A antibody may be viewed as an ecological determinant in the oral cavity by affecting oral microorganisms and possibly their by-products.
Resistance of
Salmonella typhimurium
Mutants to Galactose Death
Authors: Shuster C. W. , Rundell Kathleen
Published: January 2020

A class of galactose-resistant mutants has been derived from strains of
Salmonella typhimurium
which are defective in uridine diphosphoglucose-4-epimerase. Resistant strains are phenotypically similar to parent organisms but do not lyse in the presence of galactose. Low levels of functional epimerase can be detected in induced cells grown at 20 C but not at 37 C, and acid is not produced from galactose. Sufficient galactose is synthesized at reduced temperatures to fabricate smooth lipopolysaccharide and acceptor sites for phage P22 from galactose-deficient media. The leaky nature of these mutants may account for resistance to galactose death by maintaining galactose metabolites at a subcritical level. Glucose protects sensitive strains by control of levels of toxic metabolites by catabolite repression.
Identification of Reduced Nicotinamide Adenine Dinucleotide Phosphate-Dependent Aldehyde Reductase in A
Rhodotorula
Strain
Authors: Watson John A. , Hayashi James A. , Schuytema Eunice , Doughty Clyde C.
Published: January 2020

Reduced nicotinamide adenine dinucleotide phosphate (NADPH)-aldehyde reductase was isolated in 24% yield and 66-fold purification from a
dl
-glyceraldehyde-grown
Rhodotorula
species. This enzyme was specific for NADPH, and
d
-,
l
-, or
dl
-glyceraldehyde were equally good substrates. Other substrates had activities as follows: methylglyoxal, 50%; fructose, 33%;
d
- and
l
-arabinose, 12%;
d
-xylose, 8%;
d
-glucose, 5%;
d
-erythrose and
d
-threose, 0 to 5%. The product from the reduction of
dl
-glyceraldehyde was glycerol, as shown by high voltage electrophoresis, paper chromatography, and direct enzymatic analysis. Kinetic studies gave

K
m

values of 0.89 m
m
and 0.013 m
m
for
dl
-glyceraldehyde and NADPH, respectively. An optimal
p
H range of 6.3 to 6.7 was found for maximal activity. Reduction of NADP
+
by glycerol was not demonstrable. This
Rhodotorula
NADPH-aldehyde reductase activity was compared to similar enzymes from other sources.
Effect of R Factors and Other Plasmids on Ultraviolet Susceptibility and Host Cell Reactivation Property of
Escherichia coli
Authors: Siccardi A. G.
Published: January 2020

Some R factors, like some colicin factors, confer partial protection against the bactericidal effect of ultraviolet (UV) irradiation. Of 31 plasmids (17 R, 3
col
, and 11 R-
col
factors) tested in
Escherichia coli
K-12, 15 protected, 11 had little or no effect, and 5 caused increased UV susceptibility. The effect of representative plasmids was qualitatively the same in K-12 of wild-type UV sensitivity, λ-lysogenic or non-lysogenic, and in UV-sensitive mutants of classes
uvrA, uvrB, uvrC
, and
recA
(except that a sensitizing factor did not increase the sensitivity of two
recA
hosts). It is inferred that the UV-protecting effect of some plasmids does not result from their specifying enzymes similar to those deficient in such mutants. UV killing of multiply auxotrophic K-12, of wild-type sensitivity or
recA
or
uvrC
, was reduced by deprivation of required amino acids for 2 hr before irradiation, and further reduced if “starvation” was continued for 2 hr after irradiation. The plasmids tested in these conditions produced qualitatively the same effects as in nonstarved cells—except that in K-12 of wild-type UV sensitivity the effect of protecting plasmids was reversed (i.e. they caused decreased survival) when the cells were starved after irradiation. Two UV-protecting R factors reduced the ability of HCR
+
K-12 to support growth of irradiated phage T1.
Relationship of Footpad Sensitivity to Purified Protein Derivatives and Resistance to Airborne Infection withMycobacterium tuberculosisof Mice Vaccinated with Mycobacterial Cell Walls
Authors: Anacker R. L. , Ribi E. , Tarmina D. F. , Fadness L. , Mann R. E.
Published: January 2020

Footpads of mice sensitized by oil-treated Bacillus Calmette-Guérin (BCG) cell walls given either intravenously, subcutaneously, intradermally, intraperitoneally, or intramuscularly became swollen and reddened after injection of purified protein derivative (PPD). This reaction, greatest after intradermal and subcutaneous sensitization, generally reached a maximum about 24 hr after challenge and was still marked at 48 hr. The histological response was characterized by infiltration with both polymorphonuclear and mononuclear cells. The proportion of mononuclear cells increased with time and they predominated at 48 hr. The footpad reaction could be detected as early as 1 week after sensitization and persisted for at least 37 weeks. Footpad sensitivity to PPD and acquired resistance to airborne infection withMycobacterium tuberculosisH37Rv were correlated in that (i) both reached a peak approximately 1 month after sensitization of the mouse, and (ii) cell walls treated with NaOH or given without oil neither protected mice against challenge infection nor sensitized them to PPD. Although, as we reported previously, mice vaccinated subcutaneously or intradermally exhibited little or no enhanced resistance to experimental infection, mice given oil-treated cell walls by these routes were highly sensitive to footpad inoculation of PPD. Therefore, the footpad test cannot be used to determine immunity of the mouse to pulmonary infection with tubercle bacilli.
Differentiation Between
Clostridium sordellii
and
Clostridium bifermentans
by Gas Chromatography
Authors: Brooks J. B. , Moss C. W. , Dowell V. R.
Published: January 2020

Clostridium bifermentans
can be differentiated from
Clostridium sordellii
by gas chromatography on the basis of amines detected after growth for 6 hr in cookedmeat medium. Products detected after exposure of resting cells to amino acids gave evidence for the probable origin of the amines.
Mechanism of Tuberculostasis in Mammalian Serum II. Induction of Serum Tuberculostasis in Guinea Pigs
Authors: Kochan Ivan , Golden Carole A. , Bukovic Joann A.
Published: January 2020

The growth of tubercle bacilli in serum samples of untreated animals depends upon the availability of ionic iron which serves as a growth factor in supporting bacillary multiplication. The amount of available iron in serum is determined by the ratio between iron-saturated and iron-free transferrin; a low value for the ratio is associated with tuberculostasis (e.g., human serum, 0.4), whereas a high value is associated with the growth-supporting quality (e.g., guinea pig serum, 5.6). The treatment of guinea pigs with lipopolysaccharide ofEscherichia colior tuberculous cell wall material consistently and significantly reduced serum iron levels; a similar but less striking effect was observed in BCG-vaccinated animals. Pronounced differences were observed in the time of appearance and duration of serum hypoferremia; in lipopolysaccharide-treated animals, it appeared in 1 day and lasted for several days, whereas in BCG-vaccinated animals it appeared in about 2 weeks and lasted for much longer time periods. The induced hypoferremia was always associated with the concomitant development of serum tuberculostasis which could be neutralized by the addition of iron. These results indicate, therefore, that the mechanism of induced serum tuberculostasis in lipopolysaccharide- or tuberculous cell wall-treated and BCG-vaccinated guinea pigs is the same as that present in tuberculostatic sera of untreated animals.
Periplasmic Structure of Frozen-Etched and Negatively Stained Cells of
Bacillus licheniformis
as Correlated with Penicillinase Formation
Authors: Ghosh B. K. , Lampen J. O. , Remsen C. C.
Published: January 2020

Bacillus licheniformis
strain 749/C (constitutive for penicillinase formation) and uninduced cells of strain 749 (penicillinase-inducible) were examined after freezeetching. In the early stationary phase, strain 749/C organisms had clusters of vesicles (30 to 40 nm in diameter) on the outer surface of the plasma membrane. These are randomly distributed on the membrane, including the region of septum formation. The vesicles are not intimately associated with the plasma membrane, and their inner and outer surfaces are devoid of particles. Periplasmic vesicles were not detected by freeze-etching in strain 749 (uninduced) or in young cells of 749/C; however, the membrane of mid-logarithmic phase 749/C cells had a corrugated appearance. Negatively stained 749/C cells (logarithmic phase) also showed many vesicular and tubular bodies in the periplasm as well as septal and cytoplasmic mesosomes of typical morphology. The periplasmic structures appear to be formed either by evagination of plasma membrane or by migration of vesicular bodies from the membranous pockets of the cytoplasm. Stationary phase cells of 749/C still have many periplasmic vesicular bodies; however, the mesosomes are greatly reduced both in number and size. In sharp contrast, strain 749 organisms have very few structures similar to the periplasmic bodies of strain 749/C. These findings support our previous view that penicillinase-producing cells of 749/C have periplasmic membranous structures that are rare in the uninduced strain 749, though there is some lack of correspondence between freeze-etching, negative staining, and thin section data. These structures may be important for the retention or storage of penicillinase in the cell.
Comparative Study of the Structure of Gas Vacuoles
Authors: Cohen-Bazire Germaine , Kunisawa Riyo , Pfennig Norbert
Published: January 2020

The fine structure of gas vacuoles was examined in two blue-green algae, two green bacteria, three purple sulfur bacteria, and two halobacteria. The gas vacuole is a compound organelle, composed of a variable number of gas vesicles. These are closed, cylindrical, gas-containing structures with conical ends, about 80 to 100 nm in width and of variable length, ranging from 0.2 to over 1.0 μm. The wall of the gas vesicle is a non-unit membrane 2 to 3 nm in thickness, bearing very regular striations with a periodicity of 4 nm, oriented more or less at right angles to the long axis of the cylinder. This fine structure could be clearly resolved in isolated gas vesicles prepared from a blue-green alga and from
Halobacterium halobium
, and its presence in the gas vesicles of the green bacterium
Pelodictyon clathratiforme
was inferred from thin sections. The gas vacuole thus appears to be a homologous organelle in all of these procaryotic groups. Minor differences with respect to the length and arrangement of the gas vesicles were observed. In blue-green algae and green bacteria, the vesicles are relatively long and tend to be arrayed in parallel bundles; in purple sulfur bacteria and
Halobacterium
, they are shorter and more irregularly distributed in the cell.
Relationship between the Latent Form and the Active Form of the Autolytic Enzyme of
Streptococcus faecalis
Authors: Pooley Harold M. , Shockman Gerald D.
Published: January 2020

A 10-hr starvation of
Streptococcus faecalis
ATCC 9790 for the amino acids methionine and threonine results in cells which are resistant to autolysis and which contain greatly reduced quantities of both active and latent (proteinase activable) forms of the autolytic enzyme (an
N
-acetyl-muramide glycanhydrolase). Cell walls were isolated from cells harvested at various times during the recovery from such starvation and were assayed for active and latent forms of the autolysin. Within 10 min of recovery the latent enzyme began to increase. Only after 30 to 60 min did the active enzyme begin to increase; after a similar lag, the cells' proneness to lysis markedly increased. The intracellular localization of both forms of the autolysin was examined, using as an experimental tool the ability of added cell wall to bind autolysin.
14
C-lysine-labeled, inactivated cell walls were added to exponential-phase cells, which were then disrupted, and the mixed wall population was isolated. Measurement of the
14
C release during wall autolysis indicated that the active enzyme in the cells was not available for binding to the added
14
C-labeled walls and was therefore wall-bound in vivo. In contrast, up to 85% of latent autolysin activity was found to have been efficiently bound to the added
14
C walls. The results obtained suggest (i) cellular autolysis is a reflection of the level of active enzyme and not of latent enzyme, and (ii) autolysin is synthesized and mainly located in the cytoplasm as an inactive latent precursor (proenzyme) which is transported to sites on the cell wall associated with wall biosynthesis, where it becomes activated.
Identification of
Bacillus subtilis
NRRL B-3275 as a Strain of
Bacillus pumilus
Authors: Lovett Paul S. , Young Frank E.
Published: January 2020

The physiological and biochemical properties of a species of
Bacillus
previously identified as
B. subtilis
NRRL B-3275 (B-3275) were compared with those of seven strains of
B. pumilus
and five strains of
B. subtilis
. The biotin requirement of B-3275, its inability to hydrolyze starch, and its failure to reduce nitrate indicate that the organism is more closely related to the
B. pumilus
strains than to those of
B. subtilis
. Hybridization of deoxyribonucleic acid (DNA) from B-3275 with that of the strains of
B. pumilus
showed a binding efficiency (compared with the homologous reaction) of 58 to 99%, depending on the strain. Hybridization with the DNA from any of the strains of
B. subtilis
did not exceed 24%. DNA from B-3275 was unable to transform two amino acid auxotrophic markers to prototrophy in a highly competent strain of
B. subtilis
168. We conclude that B-3275 is a strain of
B. pumilus
which we designate as
B. pumilus
NRRL B-3275.
Biochemical Studies of Bacterial Sporulation and Germination XVII. Sulfhydryl and Disulfide Levels in Dormancy and Germination
Authors: Setlow Peter , Kornberg Arthur
Published: January 2020

A fourfold increase in sulfhydryl content upon germination of
Bacillus megaterium
spores was observed by the standard fluorescein mercuric acetate assay as reported by others. However, assay of ruptured dormant spores or the use of
N
-ethylmaleimide and a denaturing agent on intact spores showed a constant sulfhydryl level in dormancy and in germination. The apparent increase in sulfhydryl groups observed on germination was shown to be due to inaccessibility of most sulfhydryl groups in the dormant spore to sulfhydryl reagents. The disulfide content of dormant spores showed no change on germination, nor was any evidence found for production of low-molecular-weight sulfhydryl or disulfide compounds during germination.
Regulation of the Formation of Acid Phosphatases by Inorganic Phosphate in
Aspergillus ficuum
Authors: Shieh T. R. , Wodzinski R. J. , Ware J. H.
Published: January 2020

Two types of extracellular acid phosphatases are synthesized by
Aspergillus ficuum
NRRL 3135: a nonspecific orthophosphoric monoester phosphohydrolase (EC 3.1.3.2) with an optimum
p
H of 2.0, and an enzyme with restricted specificity, a mesoinositol-hexaphosphate phosphohydrolase (EC 3.1.3.8; phytase) with an optimum
p
H of 5.5. Although the
p
H 5.5 enzyme is termed a phytase, both enzymes hydrolyze phytin. Synthesis of the enzymes is repressed by high orthophosphate concentrations in the fermentation medium. The highest total level for each enzyme is synthesized in low orthophosphate medium. In high orthophosphate medium, more
p
H 5.5 enzyme is produced than
p
H 2.0 enzyme. In low orthophosphate medium, more
p
H 5.5 enzyme is produced than
p
H 2.0 enzyme during the early stages of growth, but the reverse occurs after 5 days. The enzymes are differentiated by heat denaturation at acid and alkaline
p
H levels. They are separated into two distinct fractions on Sephadex G-100 followed by carboxymethylcellulose column chromatography. This indicates that the two enzymes are structurally different. The
K
m
for both enzymes is 1.25 m
m
when calcium phytate is the substrate. Orthophosphate competitively inhibits the
p
H 2.0 (
K
i
= 1.1 × 10
−2
m
) but not the
p
H 5.5 phosphatase. Neither enzyme is denatured by 50% (w/v) urea or inhibited by 0.01
m
tartrate. Thus, they differ from human prostatic phosphatase.
Metabolism of
Bacillus thuringiensis
in Relation to Spore and Crystal Formation
Authors: Yousten A. A. , Rogoff M. H.
Published: January 2020

A general pattern of metabolism was determined for
Bacillus thuringiensis
grown in a glucose-yeast extract-salts medium. The pattern did not differ significantly from that of
B. cereus
grown in a similar medium. Acetic acid produced from glucose during exponential growth was further catabolized in the early sporulation phase of growth, at which time the specific activity of aconitate hydratase increased markedly. Fluoroacetate and α-picolinate prevented the removal of accumulated acid, and the resulting low
p
H inhibited spore and crystal synthesis. Neither crystal-related antigens nor insect toxicity was shown by cells whose crystal synthesis was inhibited in this way. α-Picolinate prevented the normal increase in specific activity of aconitate hydratase without inhibiting exponential growth. It also inhibited aconitate hydratase in vitro, but only if preincubated with the enzyme. α-Picolinate did not inhibit the increase in specific activity of aconitate hydratase or spore and crystal synthesis in a medium buffered near neutrality. Chloramphenicol and actinomycin D inhibited crystal enlargement and sporulation when added to cells in which small crystals had already begun to form. Typical messenger ribonucleic acid-dependent protein synthesis, rather than the type associated with peptide antibiotic synthesis, is thus indicated for the synthesis of crystal peptide subunits.
Microgermination of
Bacillus cereus
Spores
Authors: Hashimoto Tadayo , Frieben W. R. , Conti S. F.
Published: January 2020

The biphasic nature of germination curves of individual
Bacillus cereus
T spores was further characterized by assessing the effects of temperature, concentration of germinants, and some inorganic cations on microgermination. Temperature was shown to affect both phases of microgermination as well as the microlag period, whereas the concentration of
l
-alanine and supplementation with adenosine exerted a significant effect only on the microlag period. The germination curves of individual spores induced by inosine were also biphasic and resembled those of spores induced by
l
-alanine. High concentrations (0.1
m
or higher) of calcium and other inorganic cations prolonged both phases of microgermination, particularly the second phase, and had a less pronounced effect on the microlag period. The second phase of microgermination was completely inhibited when spores were germinated either in the presence of 0.3
m
CaCl
2
or at a temperature of 43 C; this inhibition was reversible. Observations on the germination of spore suspensions (kinetics of the release of dipicolinic acid and mucopeptides, loss of heat resistance, increase in stainability, decrease in turbidity and refractility) were interpreted on the basis of the biphasic nature of microgermination. Dye uptake by individual spores during germination appeared also to be a biphasic process.
Phosphorylation of Methyl-α-
d
-Glucopyranoside in Polymyxin B-Treated
Salmonella typhimurium
Authors: Teuber Michael
Published: January 2020

Phosphoenolpyruvate-dependent phosphorylation of methyl-α-
d
-glucopyranoside in
Salmonella typhimurium
is increased by the membrane active polypeptide antibiotic polymyxin B whereas active transport ability is abolished.
Nonoxidative Pentose Phosphate Pathway in
Veillonella alcalescens
Authors: Michaud R. N. , Carrow J. A. , Delwiche E. A.
Published: January 2020

Crude cell-free extracts of
Veillonella alcalescens
C1, an anaerobe unable to ferment glucose, were assayed for individual enzymes of the pentose phosphate pathway. Glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities were not detectable. Constituent enzymes of the nonoxidative limb of the pentose phosphate pathway were demonstrable. The presence of transaldolase, transketolase, phosphoribose isomerase, and phosphoribulose epimerase in this organism suggests a primarily biosynthetic role for these enzymes. It is postulated that ribose is synthesized from lactate in
V. alcalescens
C1 via a modified reversal of glycolysis and the nonoxidative limb of the pentose phosphate pathway.
Chemical Composition of Exosporium from Spores of
Bacillus cereus
Authors: Matz L. L. , Beaman T. Cabrera , Gerhardt Philipp
Published: January 2020

Homogeneous fragments of exosporium were extricated in centigram amounts from dormant spores of
Bacillus cereus
and analyzed for intrinsic constituents. The membrane proved to be chemically complex but not unique, consisting mainly of protein (52%), amino and neutral polysaccharides (20%), lipids (18%), and ash (4%). Seventeen common amino acids were identified by chromatography, and were present in usual proportions except for low levels of cystine-cysteine, methionine, tyrosine, and histidine. Glucosamine was the only amino sugar, and glucose and rhamnose were the principal neutral sugars. The lipid fraction contained 5.5% cardiolipin as the only phospholipid, 12.5% neutral lipids, and at least 19 fatty acids, among which normal C
16
and C
18
ones predominated. Calcium and phosphorus occurred in the ash. Small amounts of teichoic, ribonucleic, and dipicolinic acids were believed to represent contamination.
Morphogenesis and Fine Structure of
Leucothrix mucor
and Effects of Calcium Deficiency
Authors: Snellen James E. , Raj H. D.
Published: January 2020

Phenotypic expressions of morphogenesis and fine structure of
Leucothrix mucor
were determined when the organism was grown with and without added CaCl
2
in a synthetic seawater medium. Evidence is presented to show that a bulb can form in the absence of a knot formation and that a bulb may give rise to a “germ-tube.” In comparison with normal cells, which show transverse septa at right angles to the axis in dividing cells, the bulbs exhibited transverse septa at odd angles, which may explain the mechanism of bulb formation. The most striking morphological feature due to Ca
++
deficiency was the absence of rosettes; instead, the culture showed an extremely filamentous morphology and a peculiar cord formation. Also, the Ca
++
-deficient cells contained heavily stained intracytoplasmic granules which possibly represent tight packing of the smaller particles of ribonucleoprotein. Various bulbous forms observed in the Ca
++
-deficient culture showed more pronounced elaboration of mesosomes as intracytoplasmic structures than those seen in the complete medium.
Proposal for Classifying Strain PG-24 and Related Canine Mycoplasmas asMycoplasma edwardiisp. n
Authors: Tully Joseph G. , Barile Michael F. , Del Giudice Richard A. , Carski Theodore R. , Armstrong Donald , Razin Shmuel
Published: January 2020

Mycoplasmas recovered recently from dogs were found unrelated to three classified canineMycoplasmaserotypes but were similar in biological and serological properties to aMycoplasmastrain (C21, PG-24) isolated 18 years earlier. It is proposed that strains with the characteristics described be designatedMycoplasma edwardiisp. n.
Chromosomal Location of a Gene Defining Nicotinamide Deamidase in
Escherichia coli
Authors: Dickinson Ellen S. , Sundaram T. K.
Published: January 2020

A gene specifying the enzyme nicotinamide deamidase has been mapped, by the interrupted mating technique, at about 33 min on the chromosome of
Escherichia coli
K-12.
Effect of the Thiol-Oxidizing Agent Diamide on the Growth of
Escherichia coli
Authors: Wax Richard , Rosenberg Eugene , Kosower Nechama S. , Kosower Edward M.
Published: January 2020

Oxidation of glutathione within
Escherichia coli
cells by diamide, (CH
3
)
2
NCON=NCON(CH
3
)
2
, stops growth but does not cause cell death. Normal growth rates are resumed after periods which vary in length according to the diamide concentration. Consumption of excess reagent with added glutathione quickly reverses the inhibition. Another thiol-oxidizing agent, azoester, C
6
H
5
N=NCOOCH
3
, causes lysis.
Carbon Dioxide Fixation by Extracts of
Streptococcus faecalis
var.
liquefaciens
Authors: Hartman Ronald E.
Published: January 2020

Extracts of cells of
Streptococcus faecalis
var.
liquefaciens
strain 31 incorporated
14
CO
2
into aspartate. Dialyzed extracts produced radioactive oxalacetate in the absence of exogenously added glutamate and pyridoxal-5′-phosphate and produced radioactive aspartate in the presence of these components. Reduced nicotinamide adenine dinucleotide or reduced nicotinamide adenine dinucleotide phosphate could not be substituted for adenosine triphosphate (ATP); phosphoenolpyruvate even in the presence of nucleoside diphosphates could not replace pyruvate plus ATP; propionate plus coenzyme A (CoA) could not replace pyruvate in supporting CO
2
fixation by cell extracts. Fixation by dialyzed cell extracts required pyruvate, ATP, MgSO
4
, and was stimulated by biotin, KCl, 2-mercaptoethanol, CoA, and acetyl CoA. Inhibition of fixation occurred when avidin, NaCl, oxalacetate, or aspartate was added to dialyzed extracts. On the basis of the products formed and the effects of substrates and cofactors on the fixation reaction, it was concluded that pyruvate carboxylase is responsible for CO
2
fixation in this microorganism.
Formate as an Intermediate in the Bovine Rumen Fermentation
Authors: Hungate R. E. , Smith W. , Bauchop T. , Yu Ida , Rabinowitz J. C.
Published: January 2020

An average of 11 (range, 2 to 47) μmoles of formate per g per hr was produced and used in whole bovine rumen contents incubated in vitro, as calculated from the product of the specific turnover rate constant,
k
, times the concentration of intercellular formate. The latter varied between 5 and 26 (average, 12) nmoles/g. The concentration of formate in the total rumen contents was as much as 1,000 times greater, presumably owing to formate within the microbial cells. The concentration of formate in rumen contents minus most of the plant solids was varied, and from the rates of methanogenesis the Michaelis constant,
K
m
, for formate conversion to CH
4
was estimated at 30 nmoles/g. Also, the dissolved H
2
was measured in relation to methane production, and a
K
m
of 1 nmole/g was obtained. A pure culture of
Methanobacterium ruminantium
showed a
K
m
of 1 nmole of H
2
/g, but the
K
m
for formate was much higher than the 30 nmoles for the rumen contents. It is concluded that nonmethanogenic microbes metabolize intercellular formate in the rumen. CO
2
and H
2
are the principal substrates for rumen methanogenesis. Eighteen per cent of the rumen methane is derived from formate, as calculated from the intercellular concentration of hydrogen and formate in the rumen, the Michaelis constants for conversion of these substrates by rumen liquid, and the relative capacities of whole rumen contents to ferment these substrates.
Hemin-Deficient Mutants of
Salmonella typhimurium
Authors: Săsărman A. , Sanderson K. E. , Surdeanu M. , Sonea S.
Published: January 2020

Nine hemin-deficient mutants of
Salmonella typhimurium
LT2 were isolated as neomycin-resistant colonies. Five of these mutants could be stimulated by Δ-aminolevulinic acid (Δ-ALA), thus representing
hemA
mutants. Since
S. typhimurium
LT2 is not able to incorporate hemin, the identification of the mutants not stimulated by Δ-ALA was made on the basis of the simultaneous loss of catalase activity and cytochromes. The
hemA
gene was mapped by conjugation in the
trp
region, probably in the order
purB-pyrD-hemA-trp;
the episome FT
71
trp
does not carry the
hemA
gene. Transductional intercrosses by phage P22 indicate that
hemA 11, 12, 13,
and
37
are at very closely linked sites, whereas
hemA14
is at a more distant site in the same or an adjacent gene. No joint transduction was detected between
hemA
and
trp
or
pyrF
. The loci affected in the other hemin-deficient mutants were linked in conjugation to the
pro
+
marker (frequency of linkage, 88 to 97%), but cotransduction of the two markers could not be obtained. The episome F
lac hem purE
, which originates from
Escherichia coli
K-12, could complement these hemin-deficient mutants of
S. typhimurium
LT2. As a result, the sequence of the markers on the chromosome of
S. typhimurium
LT2 is probably
pro heme purE
, analogous to the sequence found in
E. coli
K-12. Thus, the chromosome of
S. typhimurium
also possesses two
hem
regions, with a location similar to that described in
E. coli
K-12.