bluuebunny/crossref_metadata_2025 huggingface dataset:
Group B Arbovirus Structural and Nonstructural Antigens I. Serological Identification of Saint Louis Encephalitis Virus Soluble Antigens
Authors: Qureshi Ateef A. , Trent Dennis W.
Published: January 2020

Saint Louis encephalitis virus-infected cells were solubilized with Brij-58 nonionic detergent, and the clarified supernatant fluid was analyzed by diethyl-aminoethyl-cellulose chromatography. Three serologically distinguishable antigens were identified in viral protein peaks which eluted from diethylaminoethyl-cellulose at 0.05, 0.075, 0.125 and 0.2
m
KCl. Antigen I was found in all column peaks, whereas antigen II was found only in the 0.05
m
KCl eluate and antigen III was found only in the 0.2
m
eluate. Immunodiffusion studies revealed that antigens I and II share at least one antigenic determinant and that antigen III is immunologically distinct from either antigen I or II. Polyacrylamide gel electrophoresis of
3
H-leucine-labeled concentrated column peaks revealed that antigens I and II contain the major viral structural envelope and core proteins, respectively. Antigen III contains the largest virus-induced nonstructural polypeptide observed by polyacrylamide gel electrophoresis of
3
H-leucine-labeled Saint Louis encephalitis virus-infected cells.
Rapid Batch Method for the Preparation of Fluorescein Isothiocyanate-Conjugated Antibody
Authors: Calcagno John V. , Sweeney Michael J. , Oels Helen C.
Published: January 2020

A rapid, reproducible, and relatively simple method for the preparation of large quantities of fluorescein isothiocyanate-conjugated antibody is presented. Tagged globulins with fluorochrome-protein ratios of 1 and 2 were eluted from diethylaminoethyl-cellulose in batch form by using sodium phosphate-buffered 0.1 M and 0.2 M saline, respectively. The intensity and specificity of the fluorescence due to this antibody was quite high, as determined by the ability of untagged antibody to block the immunological reactivity of the fluorescent antibody and by the lack of fluorescence of antigenically unrelated material when treated under identical conditions.
Neutralization of the Phagocytosis-Enhancing Acitivity of Interferon Preparations by Anti-Interferon Serum
Authors: Donahoe Robert M. , Huang Kun-Yen
Published: January 2020

A wide variety of distinctive mouse interferon (IF)-containing preparations consistently exhibited the phagocytosis-enhancing activity reported previously. The enhancing as well as the antiviral activity of IF preparations were neutralized by a highly specific anti-IF serum.
Immunity to
Salmonella typhimurium
Infection: Characterization of Antigens in Active Protection by Polyacrylamide Gel Electrophoresis
Authors: Houchens David P. , Wright George L.
Published: January 2020

Immunity to challenge with
Salmonella typhimurium
(strain SR-11) was studied in NIH/NMRI Swiss mice. Viable organisms induced the best protection, but ribosomes were the best immunogen of the subcellular fractions. Antigens with lower concentrations of protein did not induce protection as well as antigens which had higher concentrations of protein. Acrylamide gel electrophoresis of the SR-11 subcellular components, fractionation, and differential staining of the gel columns indicated that the immunogen is a high-molecular-weight glycoprotein or mucopolysaccharide.
Degradation of Coxsackievirus Type A9 by Proteolytic Enzymes
Authors: Herrmann John E. , Cliver D. O.
Published: January 2020

The means by which coxsackievirus type A9 (CA9) is inactivated by proteolytic enzymes was investigated. After reaction of
14
C-leucine-labeled CA9 with Pronase, free leucine was liberated as measured by radiochromatography. Treatment of
14
C-leucine-labeled CA9 with trypsin or proteolytic filtrates of
Pseudomonas aeruginosa
caused the release of a variety of labeled substances. The extent of viral ribonucleic acid (RNA) release after exposure of CA9 to Pronase was determined by RNA infectivity tests or trichloroacetic acid solubility tests. Infective viral RNA was found not to be consistently released by reaction of CA9 with Pronase, but further treatment with 1% sodium dodecyl sulfate at pH 7.0 promoted viral RNA release. Sodium dodecyl sulfate treatment of CA9 that had not been reacted with Pronase did not inactivate virus or cause viral RNA release. Reaction of Pronase with
32
P-labeled CA9 resulted in the liberation of virus components soluble in cold trichloroacetic acid, whereas untreated CA9 or CA9 reacted with ribonuclease were precipitated by cold trichloroacetic acid. These results demonstrate that the primary means by which protease-sensitive enteroviruses are inactivated is by degradation of the virus capsid, with subsequent release of viral RNA.
Extracellular Esterases of Group A Streptococci
Authors: Hayano Seiki , Tanaka Atsuko
Published: January 2020

Methods were devised to prepare and estimate quantitatively the extracellular esterases of group A streptococci (STE). Two types of esterase preparations were prepared, corresponding to Stock and Lynn's serotype I (STE I) and II (STE II). The former was prepared from
Streptococcus pyogenes
strain SS379 (group A, type 40), and the latter from
S. pyogenes
strain 69882 (group A, type 49). Effects of cupric acetate, sera of various animals, and rabbit antisera to STE I and II on the activities of STE were tested. Cupric acetate added in substrate solution inhibited the activities of STE according to the concentration of cupric acetate. Sera of various animals (calf, guinea pig, horse, sheep, and pig) stimulated the activities of STE and protected them from the inhibitory effect of cupric acetate. Among the sera of various animals, horse serum gave the most potent activating and protecting effect, and was the serum from which the effective factor (horse serum-activating factor, HAF) was extracted by precipitation with ammonium sulfate. STE I was not stimulated by serum or HAF as markedly as STE II, but both enzymes were protected likewise by serum or HAF from the inhibition of cupric acetate. Antibodies to STE I and II were prepared in rabbits. Their effects on STE resembled that of HAF, but differed from the latter in immunological specificity. Activities of STE were stimulated and protected more markedly by homologous antiserum than by heterologous antiserum or normal rabbit serum. Applying suitable conditions, we defined the potency of STE and differentiated antibodies to STE I and II and normal serum quantitatively.
Relationship of Adenovirus to Lymphocytes in Naturally Infected Human Tonsils and Adenoids
Authors: Van Der Veen J. , Lambriex M.
Published: January 2020

Purified lymphocytes from human tonsil and adenoid specimens were cultured with and without phytohemagglutinin. Adenovirus was isolated from lymphocytes of 8 of 90 specimens tested. With one exception, it was necessary to culture the lymphocytes before infectious virus could be detected. Phytohemagglutinin stimulation enhanced the recovery of virus. The results suggest that lymphocytes in tonsils and adenoids may be naturally infected with adenovirus and that, in positive cultures, at least 1 of every 10
7
cells harbors virus or viral precursor at initiation of the cultures. Adenovirus was demonstrated directly in fresh suspensions of unpurified cells from tonsils and adenoids in seven cases. In five of these cases, at least 1 of every 10
6
cells contained infectious virus. Adenovirus was isolated from 61 (62%) of 98 tonsil and adenoid specimens by the conventional method of tissue fragment culture after various periods of cultivation. The viruses isolated were of serotypes 1, 2, 5, and 6.
Granulomatous Hypersensitivity to Trehalose-6,6′-Dimycolate (Cord Factor) in Mice Infected with BCG
Authors: Bekierkunst A. , Yarkoni E.
Published: January 2020

Cord factor in the form of emulsion is unable to sensitize mice to react with a more extensive granulomatous response to a subsequent challenge with the same substance. Mice infected with BCG bacilli are sensitized to cord factor. Such animals react to administration of cord factor with a very extensive granulomatous response, much stronger than normal ones. This hypersensitivity seems to be specific and is distinct from delayed hypersensitivity to PPD. PPD, administered to mice sensitized with BCG bacilli, has no effect on the granulomatous response. Our findings and their importance are discussed.
Comparison of the Capillary Tube and Explant Methods for the In Vitro Assay of Tuberculin
Authors: Heilman Dorothy H. , Affronti Lewis F. , Leichner John P.
Published: January 2020

Purified antigens prepared from mycobacteria were tested for nonspecific toxicity and tuberculin activity by two in vitro methods. One technique utilized measurements of macrophage migration in 4-day-old cultures of spleen from normal and tuberculin-sensitive rabbits. The other method was a modification of the capillary tube technique. To obtain enough peritoneal macrophages for good quantitation, changes were made in the method of harvesting cells, and in some instances exudates from two or more Wright strain no. 13 guinea pigs were combined. The capillary tube method was as sensitive as the explant method for detecting nonspecific toxicity. Each tuberculin assay included a group of control cultures of cells from sensitive animals, test groups containing two widely spaced concentrations of a standard tuberculin, PPD-S, and one or more concentration of the tuberculin to be tested. Macrophages from tuberculin-sensitive animals were regularly inhibited by 0.25 μg of PPD-S per ml with both in vitro methods. The potency of the test tuberculins relative to that of PPD-S was somewhat greater in capillary tube assays than in explant cultures. A purified tuberculopolysaccharide was equally inhibitory for both normal and tuberculin-sensitive cells in both culture systems.
Induction of Interferon in L Cells by Reoviruses
Authors: Gauntt C. J.
Published: January 2020

L cell cultures challenged with reoviruses types 1 and 3 produced little to no detectable interferon under conditions which permitted virus replication and under conditions which prevented replication of a temperature-sensitive mutant strain of reovirus type 3 (ts-1). Ultraviolet-irradiated reoviruses and double-stranded ribonucleic acids extracted from purified reovirus type 3 also induced little to no interferon in L cell cultures. Under similar conditions, MM virus proved to be an effective inducer of interferon. Exposure of L cells to interferon prior to challenge with virus (priming) failed to enhance interferon production upon subsequent challenge with reovirus although priming increased the amount of interferon produced following MM virus challenge. L cell cultures that were challenged with reovirus type 3 and subsequently with either MM or Colorado tick fever viruses produced similar titers of interferon as cell cultures that were challenged with either MM or Colorado tick fever virus alone, respectively. These data show that the presence of production of reovirus type 3 double-stranded ribonucleic acid is not sufficient for induction of interferon in L cell cultures and that additional processes which are required for induction of interferon in L cell cultures are not expressed by reovirus type 3.
Physical Separation of Streptococcal Nicotinamide Adenine Dinucleotide Glycohydrolase from Streptolysin O
Authors: Shany S. , Grushoff Phyllis S. , Bernheimer Alan W.
Published: January 2020

Streptococcal nicotinamide adenine dinucleotide glycohydrolase (NADase) with a molecular weight of about 55,000 and an isoelectric pH of 8.55 was isolated from crude streptolysin O (SLO) preparations. NADase differed from SLO in size, charge, and immunological behavior. Streptococcal NADase is considered to have no role in the hemolytic process because it has no hemolytic activity; conversely, partially purified SLO showed no NADase activity. The hemolytic activity of crude SLO was completely inhibited by anti-tetanolysin, whereas the NADase activity in the same reaction mixture was unaffected. Experiments involving double diffusion in agar also demonstrated immunological nonidentity of the two proteins.
Extracellular Antigens from
Listeria monocytogenes
II. Cytotoxicity of Hemolytic and Lipolytic Antigens of
Listeria
for Cultured Mouse Macrophages
Authors: Watson B. B. , Lavizzo Jewel C.
Published: January 2020

Purified preparations of hemolytic and lipolytic antigens were shown to be cytotoxic for tissue cultures of mouse peritoneal macrophage monolayers. The percent reduction in viability was proportional to the concentration of antigen used although, for a brief period, the hemolytic antigen stimulated the growth of cultured macrophages. Hemolytic and lipolytic antigens were synthesized by an hemolysin-producing strain (7973), but were deficient or totally lacking in nonhemolysin-producing strains (9037-7, 4219). The inoculation of strain 7973 onto macrophage monolayers at a bacterium to macrophage ratio of 10:1 resulted in a complete loss of viable phagocytic cells. The decrease in viable phagocytic cells was accompanied by an increase in viable listeria. Pronounced cytopathic activity with proliferation of listeria was also observed when 200 U of lipolytic antigen were added to macrophage monolayers that had been infected with the smooth type nonhemolysin-producing strain (4219) on the previous day. On the other hand, a high percentage of macrophages survived when exposed to nonhemolysin-producing strains only. In this case, listeria were not recovered by routine culture methods. The data presented do not establish that phagocytic cell destruction with accompanying microbial proliferation is the direct result of hemolysin elaboration; however, they suggest that there is need for further exploration of the role of the hemolysin in listeriosis.
Differential Medium for
Vibrio cholerae
Authors: Schrank Gordon D. , Stager Charles E. , Verwey W. F.
Published: January 2020

A differential medium designed for rapid presumptive identification of
Vibrio cholerae
was described and shown to be useful for enumeration of viable cholera vibrios in the presence of other intestinal bacteria.
Preparation and Biological Properties of Water-Soluble Adjuvant Fractions from Delipidated Cells of
Mycobacterium smegmatis
and
Nocardia opaca
Authors: Adam A. , Ciorbaru R. , Petit J. F. , Lederer E. , Chedid L. , Lamensans A. , Parant F. , Parant M. , Rosselet J. P. , Berger F. M.
Published: January 2020

Digestion by lysozyme of delipidated cells of
Mycobacterium smegmatis
liberates a water-soluble immunoadjuvant fraction which is chemically very similar to the water-soluble adjuvant (WSA) obtained previously from purified cell walls, but which contains somewhat more non-peptidoglycan amino acids. The yield of peptidoglycan-arabinogalactan complex is about 10 times greater starting from whole cells than from cell walls. The main biological properties of this “neo-WSA” are described: it increases circulating antibodies to ovalbumin in guinea pigs, it does not produce polyarthritis in rats or induce hypersensitivity to tuberculin, it does not increase susceptibility to histamine or hyperreactivity to endotoxin, and does not produce spleen and liver hypertrophy. Analogous immunostimulant fractions have also been obtained from delipidated cells of
Nocardia opaca
by lysozyme treatment.
New Method for the Serological Grouping of Streptococci with Specific Antibodies Adsorbed to Protein A-Containing Staphylococci
Authors: Christensen Poul , Kahlmeter Gunnar , Jonsson Svante , Kronvall Göran
Published: January 2020

A coagglutinating reagent has recently been developed and utilized in capsule typing of pneumococci. The reagent consists of stabilized staphylococci coated with specific antibody via the gamma globulin Fc-protein A reaction thereby orienting the antibody molecules with the antigen-reactive Fab parts outwards. Reagent staphylococci coated with antibodies directed against streptococcal group-specific antigens have been used in the present investigations. Overnight cultures of streptococci were treated with trypsin and then directly tested for coagglutination of selected reagent staphylococci on glass slides. In all, 179 strains of streptococci were analyzed by the method described. The results were compared with grouping by using Lancefield extracts in counter-immunoelectrophoresis and were shown to agree completely. The coagglutination method described proved to be accurate, rapid, and simple.
Growth Requirements of Pathogenic Leptospira
Authors: Staneck Joseph L. , Henneberry Richard C. , Cox C. D.
Published: January 2020

Nutritional requirements for growth at 30 C of
Leptospira pomona
and
L. canicola
have been determined. Both pathogenic serotypes initially required bovine serum albumin (BSA) for growth in a medium (SM-4) which permitted growth of the water isolate B-16. Requirement for BSA was eliminated by (i) removing much of the apparent toxicity of free fatty acids in Tween 80 on an anion exchange column, (ii) decreasing extended lag periods observed from small inocula by incorporation of pyruvate into the medium, (iii) the addition of acetate to permit full utilization of substrate fatty acids in Tween 80, and (iv) the addition of glycerol to decrease generation times. Physiologic significance of these findings is discussed, and the possibility is suggested that apparent toxicity of fatty acids for leptospires may result from their auto-oxidation products. The resulting protein-free medium (SM-5) permitted the growth of pathogens at 30 C to high cell yields in low inocula. Highly virulent and avirulent strains from the same clone of
L. canicola
Moulton were used to determine additional growth requirements associated with virulence. As incubation temperatures were increased from 30 C to those of mammalian hosts, virulent cells required biotin at 35 C and higher levels of K
+
and Mg
2+
at 37 C. Additional Fe
2+
eliminated the necessity for removing the toxicity of Tween 80 by anion exchange. Significance of these physiologic studies are discussed in relationship to virulence. The final protein-free medium (SM-6) grew highly virulent
L. canicola
from tissue to high yields from low inocula at 37 C with no loss in virulence over several transfers.
Histamine Release Induced by Human Leukocyte Lysates: Effect of Metabolic Inhibitors and Carbohydrates
Authors: Kelly Michael T. , White Arthur
Published: January 2020

The effects of metabolic inhibitors and carbohydrates on histamine release induced by human leukocyte lysates have been studied to gain an understanding of the mechanism of histamine release in this system. Dinitrophenol produced 80% inhibition of histamine release at a concentration of 10
−3
M. Iodoacetic acid inhibited release to a lesser extent, and deoxyglucose had relatively little effect. Colchicine was a potent inhibitor of release and produced 88% inhibition at a 10
−3
M concentration. Puromycin and cycloheximide had essentially no effect on histamine release. Various sugars inhibited release with mannitol and glucose, producing greater than 50% inhibition at 10
−3
M concentrations. Other sugars inhibited histamine release to a lesser degree with the following order of potency: mannitol > glucose > sucrose > galactose > fructose. These studies suggest that intact, metabolically active cells are required for histamine release induced by human leukocyte lysates and that a secretory process is involved. The results also provide information on the possible nature of the histamine-releasing factor(s) in human leukocyte lysates.
Interaction of Gonococci with Phagocytic Leukocytes from Men and Mice
Authors: Thomas David W. , Hill James C. , Tyeryar Franklin J.
Published: January 2020

The interaction of human and mouse phagocytic leukocytes with representative virulent (F62-T1) and avirulent (F62-T4, RD-5) strains of
Neisseria gonorrhoeae
was studied in vitro. Leukocyte monolayers were incubated with gonococci for 30 min at 37 C, washed repeatedly, reincubated with fresh medium, and sampled for viable bacteria at intervals. After the initial incubation period and washing, human leukocytes retained larger numbers of viable T1 than of T4. During the subsequent 120 min of incubation, the numbers of viable T1 remained approximately constant, whereas viable counts of T4 declined by about two-thirds. In contrast, mouse leukocytes under similar conditions destroyed 70% of both types of gonococci. When human bactericidal serum was applied to infected human leukocytes, it had no effect on T4 but inactivated over 50% of T1. It is concluded that T4 are phagocytized by human leukocytes and are thus exposed to internal digestion, but are protected from bactericidal serum. T1, on the other hand, either adhere to the surface of the leukocytes or remain located so that they are neither digested by the leukocytes nor protected from bactericidal serum.
Further Studies on the Influence of Steroids on Viral Infection in Mice
Authors: Giron D. J. , Allen P. T. , Pindak F. F. , Schmidt J. P.
Published: January 2020

Certain steroids have been reported to enhance experimental viral infections, whereas others have little or no effect. Interference with the interferon system has been suggested as a possible mechanism for the viral infection-enhancing (VIE) activity of hormones. In the present study, steroids (prednisolone, progesterone, testosterone) which had no effect on MM virus infection demonstrated VIE activity against Mayaro virus infection. It is suggested that the VIE activity of a hormone may be dependent on the viral agent used for challenge. Data are also presented which suggest that, in addition to interference with the interferon system, the VIE activity of steroid hormones may be the result of at least two other actions: (i) alteration of cell membranes; (ii) stimulation of initial viral replication.
Preliminary Characterization of “Immunogenic” Ribonucleic Acid Derived from Rat Peritoneal Exudate Cells
Authors: Archer Stanley J. , Wust Carl J.
Published: January 2020

An “immunogenic” ribonucleic acid (Im-RNA) has been extracted from peritoneal exudate (PE) cells of rats that were immunized with sheep erythrocytes (SRBC). Following multiple phenol extractions and deoxyribonuclease treatment, the material obtained from PE cells was eluted from diethylaminoethyl-cellulose at 0.55 M NaCl concentration and partially purified in this procedure by a factor of 7- to 10-fold. After column chromatography, Im-RNA was found to be free of antigen based on results using
51
Cr-labeled SRBC or
14
C-dinitrophenol coupled to methylated bovine serum albumin as antigens. The Im-RNA showed a biphasic hyperchromicity curve when heated. The first phase, from 30 C to 90 C was gradual, accounting for 15.2% hyperchromicity suggestive of transfer RNA melting. No loss in immunogenic activity was observed when the Im-RNA was heated to 90 C. The second phase, from 90 C to 102 C, accounting for 15.2% further hyperchromicity, had a calculated

T
m

of 96 C. Heating above 90 C resulted in an irreversible loss of immunogenic activity. These results strongly suggested that the RNA fraction contained a highly ordered secondary structure such as might be found with double-stranded nucleic acid. The nature and function of the Im-RNA is discussed.
Pathogenesis of Aleutian Disease of Mink: Nature of the Antiglobulin Reaction and Elution of Antibody from Erythrocytes and Glomeruli of Infected Mink
Authors: Cho H. J. , Ingram D. G.
Published: January 2020

Erythrocytes from mink chronically infected with Aleutian disease virus (ADV) gave positive antiglobulin reactions with rabbit anti-mink immunoglobulin (Ig)G, anti-mink C3, and anti-mink serum, but did not react with anti-mink IgM. The strongest reaction was observed with anti-mink C3. Immunoelectrophoresis demonstrated that serum from rabbits injected with erythrocytes from ADV-infected mink gave a precipitin line with normal mink serum in the beta globulin region corresponding to C3. When normal mink erythrocytes were exposed to serum from ADV-infected mink, they were not sensitized, demonstrating that the antibodies in these mink sera were not directed against erythrocyte antigens. Glycine-hydrochloride buffer treatment of erythrocyte stromata and isolated glomeruli from ADV-infected mink yielded eluates containing serum proteins in the gamma globulin region which appeared to be IgG, and in the beta and alpha globulin regions which are probably complement components. In both erythrocyte and glomerular eluates, anti-ADV antibody was demonstrated. These findings suggested that the positive direct antiglobulin test and glomerulonephritis in Aleutian disease is due to the persistence of ADV and formation and deposition of ADV antigen-antibody-complement complexes on the erythrocyte surfaces and in glomerular capillaries.
Antimycobacterial Effect of Lysates Prepared from Immunologically Activated Macrophages
Authors: Kochan Ivan , Golden Carole A.
Published: January 2020

Lysates or heptane extracts of peritoneal (P) and alveolar (A) normal macrophages (N-M), immune macrophages (I-M), and immune-activated macrophages (IA-M) were examined for antimycobacterial activity by the agar-plate diffusion test. This test has been found suitable to reveal the antibacterial activity in 3-day incubated, but not in freshly prepared, lysates. Results showed that materials of IA-AM or I-AM and of IA-PM exerted antimycobacterial effects, whereas materials of N-PM, I-PM, and of N-AM were usually inactive. Antimycobacterial activity of lysates of AM was stronger than that of PM. The formation of antibacterial factors during an incubation of M lysates, the solubility of the factors in heptane, and various other characteristics suggested that the antimycobacterial effect was caused by the formation of toxic levels of non-esterified fatty acids. M lysates exerted equal activities against BCG, H
37
Ra, and H
37
Rv strains of tubercle bacilli. The presence of antimycobacterial activity in lysates prepared from IA-M of either BCG- or BCG-sensitized animals indicated that the potential to generate antimycobacterial activity is associated with the state of delayed hypersensitivity and the state of activation of M.
Fate of
Histoplasma capsulatum
in Guinea Pig Polymorphonuclear Leukocytes
Authors: Howard Dexter H.
Published: January 2020

Guinea pig polymorphonuclear leukocytes (PMN) were parasitized in vitro with yeast cells of
Histoplasma capsulatum.
Preparations stained after 3 h at 37 C with May Greenwald-Giemsa revealed that 87% of the yeasts were tinctorially altered. Such alterations corresponded to those displayed by fungus cells intentionally killed by heat or other means and thus the altered yeast were presumed to be dead. A combination of 10
−5
M H
2
O
2
, 10
−5
M KI, and horseradish peroxidase killed
H. capsulatum.
Death was assessed by the eosin-y dye exclusion test. All of the listed components were required for death of the fungus. A granule lysate preparation derived from guinea pig PMN leukocytes could replace the horseradish peroxidase in the fungicidal system. The granule lysates behaved in keeping with the attributes of a myeloperoxidase. Thus, PMN leukocytes and certain extracellular peroxidase systems kill the yeast cell phase of the dimorphic fungus
H. capsulatum.
Growth Curves and Clearance Rates of Virulent and Benign Venezuelan Encephalitis Viruses in Hamsters
Authors: Jahrling Peter B. , Scherer William F.
Published: January 2020

Four virulent strains of Venezuelan encephalitis virus attained higher concentrations of infectious virus in bloods of adult hamsters than two benign strains when given subcutaneously. One benign virus, the attenuated TC83 vaccine strain, reached higher concentrations in bone marrow and Peyer's patches than virulent subtype I strains, and another benign virus, subtype IV, grew to lower levels. When inoculated intracranially, the two benign viruses attained high concentrations in brain, but did not significantly alter in lethality although morbidity was increased. Intracardiac inoculation failed to increase virus concentrations of two benign viruses in blood or hematopoietic tissues above those found after subcutaneous inoculation, nor did it increase lethalities to those of virulent virus. Three benign virus strains were cleared more rapidly from hamster plasmas than six virulent strains. Differences in clearance rates were apparently not due to destruction of benign viruses by blood or tissue components. Thus viral concentrations in blood correlated directly and clearance rates inversely with hamster virulence, whereas the rate and extent of growth of a VE virus in hematopoietic or brain tissues did not correlate with ability to kill hamsters.
Prevention of Experimentally Induced Enteric Colibacillosis in Newborn Calves
Authors: Newman F. S. , Myers L. L. , Firehammer B. D. , Catlin J. E.
Published: January 2020

Severe enteric colibacillosis, characterized by profuse watery diarrhea, dehydration, apathy, hypothermia, and inability to stand, was produced in seven of eight newborn, colostrum-fed calves from nonvaccinated dams after oral challenge of calves with 10
11
viable cells of
Escherichia coli
strain B44. Twenty-nine of 32 calves from dams vaccinated with various preparations of
E. coli
strain B44 were protected against severe enteric colibacillosis after oral challenge. Calves were protected if the dams received two subcutaneous vaccinations prior to calving. Calves also were protected if the dams were given a homologous vaccine of live or killed whole cells or a broth culture supernatant vaccine via the subcutaneous and intramammary routes 1 year earlier. Nineteen of the 29 protected calves had a transient diarrhea.
Influence of Histones on the Antibody Response in Guinea Pigs to Vaccination with A/Aichi/2/68 (H3 N2) Influenza Virus
Authors: Drescher Joachim , Gillissen Günther
Published: January 2020

The influence of total, F1, and F3 histones on the primary antibody response in guinea pigs to vaccination with influenza virus A/2/Aichi/2/68 (H3 N2) was investigated. It was found that histones can greatly reduce the immunoglobulin (Ig) M and, to a smaller degree, the IgG antibody response. The immunosuppressive effect of histones was found to depend critically on the dose of virus used for vaccination, on the type and dose of histones, and on the schedule of histone application.
Distribution and Metabolism of
14
C-Tryptophan in Normal and Endotoxin-Poisoned Mice
Authors: Moon Robert J. , Tremblay Elaine S. , Morris Katherine M.
Published: January 2020

dl
-Tryptophan (benzene ring-
14
C) and its metabolites persist longer and in greater quantity in tissues of endotoxin-poisoned mice than in tissues of normal mice. Correspondingly less label is excreted in urine and feces and expired as
14
CO
2
in the poisoned animals. The distribution of label (1.1 × 10
6
dpm per microgram of tryptophan) was relatively constant whether it was administered alone or in combination with 20 mg of unlabeled
l
-tryptophan. Tryptophan must be metabolized through the tryptophan oxygenase pathway to be converted to carbon dioxide, but attempts to quantitatively correlate depressed tryptophan oxygenase activity with depressed carbon dioxide production were unsuccessful. It appears that neither tryptophan oxygenase nor substrate availability exclusively determine the quantity of tryptophan converted to
14
CO
2
except under highly selected conditions. The validity of an earlier suggestion that expired
14
CO
2
could be used to monitor in vivo tryptophan oxygenase activity is not supported by our data.
Resistance to
Mycoplasma synoviae
is Bursal Dependent
Authors: Vardaman T. H. , Landreth K. , Whatley S. , Dreesen L. J. , Glick B.
Published: January 2020

Bursectomizing small bursa line birds produced susceptibility to
Mycoplasma synoviae.
This suggested that
M. synoviae
is B cell dependent, but does not rule out a T cell effect.
Purification and Properties of the Cathepsin D Type Proteinase from Beef and Rabbit Lung and Its Identification in Macrophages
Authors: Rojas-Espinosa Oscar , Dannenberg Arthur M. , Murphy Patrick A. , Straat Patricia A. , Huang P. C. , James Stephen P.
Published: January 2020

The acid-acting proteinase, cathepsin D (EC 3.4.4.23), was purified from extracts of homogenized rabbit lung and beef lung by autolysis at acid pH, acetone and ammonium sulfate fractionation, column chromatography, and isoelectric focusing. Four isoenzymes were obtained from each source. With acid hemoglobin as the substrate, the proteinase from rabbit lung had a pH optimum of 3.0 and that from beef lung had a pH optimum of 3.6. Their activity was not affected by thiol reagents or by Fe
2+
, Mn
2+
, or Mg
2+
. One isoenzyme from rabbit lung was used to immunize a goat, and one from beef lung was used to immunize a rabbit. In immunoelectrophoresis, each resulting antiserum formed a single precipitin line with its homologous enzyme. They cross-reacted with the other three isoenzymes from the same species, but not with any isoenzyme from the other species. At high concentrations, each antiserum completely inhibited the proteolytic activity of its homologous enzyme. The antiserum against rabbit lung cathepsin D also inhibited the proteolytic activity of rabbit peritoneal and pulmonary macrophages. In limited quantities, this antiserum has now been made commercially available and is being used with labeled antibody techniques to identify under a microscope the presence of cathepsin D in macrophages and to study its role in the pathogenesis of tuberculosis.
Energetics (Adenosine 5′-Triphosphate) of
Mycobacterium lepraemurium
in Diffusion Chambers Incubated In Vitro and in Mice
Authors: Dhople Arvind M. , Hanks John H.
Published: January 2020

Adenosine 5′-triphosphate (ATP) measurements and the processing of samples have been refined to a point where the energetics and growth potential of microscopic samples of unwashed host-grown, host-dependent microbes can be investigated.
Mycobacterium lepraemurium
, the noncultivated agent of murine leprosy, was employed to examine three reports of the slow microscopic growth of this organism in the absence of host cells. A few million bacterial cells were enclosed in Rightsel- and Ito-type diffusion chambers, which were incubated in vitro and in the peritoneal cavities of mice. In the
in vitro
experiments, a complex medium containing bovine serum and mouse brain extracts, renewed three times a week, did not sustain the energetics of the bacilli. The microscopic counts declined to 72% and the ATP per culture to 9% of the original values. Very different results were obtained from chambers incubated in the peritoneal cavities of mice. The bacterial biomass increased 2.7-fold and the ATP per culture increased 2.5-fold. Because the ATP per cell was 93% of the original, this system is regarded as the first to permit the extracellular growth of a so-called “obligate intracellular microbe.” The results obtained with only 1 × 10
6
host-grown cells per assay demonstrate a significant biochemical tool for investigating the growth potential of host-grown microbes during the progression, regression, and therapy of disease.
Role of Interferon in Murine Lactic Dehydrogenase Virus Infection, In Vivo and In Vitro
Authors: Dubuy Herman , Baron Samuel , Uhlendorf Carol , Johnson Martin L.
Published: January 2020

The induction of interferon (IF) by lactic dehydrogenase virus (LDV) and the action of exogenous IF on LDV multiplication have been studied in vivo and in vitro. In vivo the induction of serum IF by LDV was shown to correspond with the virus challenge dose. In vitro LDV-infected macrophages did not produce detectable IF. As to the action of IF in vivo, the prophylactic or therapeutic administration of IF to mice reduced the LDV serum titers only temporarily. In vitro, (i) the sensitivity of LDV to IF in macrophage culture was 30 times less than that of vesicular stomatitis virus, and 5 times less than that of Sendai virus, and (ii) prolonged exposure of LDV-infected macrophages to high IF concentrations did not cure the chronic infection, but only decreased the virus titer. The probable role of IF in the establishment of the early phase of a persistent but reduced viremia in mice is discussed.
Action of Group A Streptococcus Extracellular Product(s) on the Connective Tissue of the Bovine Heart Valve
Authors: Goldstein Israël , Caravano René , Parlebas Janine
Published: January 2020

Group A streptococcal strains isolated from rheumatic fever patients were cultivated in the presence of bovine heart valves in a medium devoid of components from animal origin. Other group A streptococci and various bacteria were used as controls. The supernatant of these cultures was extracted and analyzed chemically and immunologically. The extracts prepared from cultures of two “rheumatogenic” strains in the presence of bovine heart valves showed fraction(s) containing proteins, oses (neutral and amined), and uronic and sialic acids. This fraction(s) was immunologically active with both anti-group A streptococcus and antisoluble connective glycoprotein antisera, with a partial identity reaction. Experiments with a diffusion chamber and attempts to precipitate a postulated enzyme from the culture of these strains by ammonium sulfate suggest that this action is due to an extracellular product of the bacteria. The meaning of these data in the physiopathology of rheumatic cardiac lesions is briefly discussed.
Serotyping of
Chlamydia
I. Isolates of Ovine Origin
Authors: Schachter Julius , Banks Joyce , Sugg Nancy , Sung Minnie , Storz J. , Meyer Karl F.
Published: January 2020

Eight chlamydial isolates of ovine origin were tested in a plaque reduction system using homologous and heterologous rooster antisera. The eight isolates could be separated into two separate immunotypes. Type 1 included isolates associated with ovine abortion and one agent recovered from the feces of an apparently normal sheep. Type 2 isolates were associated with polyarthritis and conjunctivitis. These two serotypes were not cross-reactive with several chlamydiae of avian origin. Further application of the plaque reduction test may provide a useful means of typing chlamydiae.
Staphylococcal Enterotoxin Synthesis During the Exponential, Transitional, and Stationary Growth Phases
Authors: Czop Joyce K. , Bergdoll Merlin S.
Published: January 2020

Small inocula (1 to 10 colony-forming units per ml of broth) of
Staphylococcus aureus
strains S-6, S-6R, and FRI-100 were employed to study growth and enterotoxin synthesis in 4% protein hydrolysate powder broths. For each strain, the exponential growth phase ended once the population approached 10
9
to 2 × 10
9
colony-forming units per ml. By that time, the concentrations of enterotoxins A and B reached the minimal level (1 to 2 μg/ml) at which the single gel diffusion tube method becomes applicable. By microslides and reverse passive hemagglutination, enterotoxins A and B were found to be synthesized during the exponential growth phase, but at different exponential rates.
Peptic Digestion of Streptococcal M Protein I. Effect of Digestion at Suboptimal pH upon the Biological and Immunochemical Properties of Purified M Protein Extracts
Authors: Cunningham Madeleine W. , Beachey Edwin H.
Published: January 2020

Purified streptococcal M protein vaccines often produce non-type-specific immunotoxic reactions in the skin or the blood of humans. In an attempt to free the type-specific M antigen (TSM) of such non-type-specific (NTSM) immunotoxic properties, purified M protein preparations were subjected to brief periods (5 to 10 min) of enzymatic digestion in dilute solutions of pepsin (20 μg/ml) at pH 5.0. Such peptic digestion abolished the ability of M protein to aggregate platelets in human platelet-rich plasma and to precipitate plasma fibrinogen. It greatly reduced (up to 16-fold) the capacity of M protein to react with the NTSM complement-fixing antibody that is normally present in human serum. In contrast, it had no effect upon the reactivity of the type-specific M antigen; the pepsin-treated M protein retained its ability to inhibit type-specific streptococcal opsonization by homologous M antibody, and, moreover, retained its ability to elicit type-specific opsonic antibody responses in rabbits.
Lipid Catabolism of Relapsing Fever Borreliae
Authors: Pickett James , Kelly Richard
Published: January 2020

Relapsing fever borreliae require lipid compounds for growth in vitro. In this study, the major pathways of lipid catabolism in three species of tick-borne relapsing fever borreliae were investigated. Thin-layer chromatography was used to compare chloroform-methanol extracts of fresh culture media with extracts of exhausted culture media after organisms were removed by centrifugation. The chromatographic data demonstrated that lysolecithin was removed from the culture media during growth of the spirochetes, whereas lecithin, sphingomyelin, triglycerides, and cholesterol esters were not affected by growth of the organisms. Sonic extracts of the organism were tested for the presence of specific enzymes of lipid catabolism. Lysolecithinase, glycerophosphorylcholine diesterase, and acid phosphatase activities were demonstrated. Thus, these organisms can sequentially dissimilate lysolecithin to fatty acids, choline, inorganic phosphate, and glycerol. Assays for phospholipases A, C, and D, α-glycerophosphate dehydrogenase, alkaline phosphatase, and lipase were negative.
Interferon Administered in the Cerebrospinal Space and Its Effect on Rabies in Rabbits
Authors: Ho Monto , Nash Carolyn , Morgan Charles W. , Armstrong John A. , Carroll Robert G. , Postic Bosko
Published: January 2020

Because combined administration of intramuscular and intravenous interferon has been partially successful in the incubationary treatment of rabies, the effect of direct interferon administration into the cerebrospinal fluid space was tested. After injecting 1,800 U of interferon into the cisterna magna or the lateral ventricle, periodic samples, obtained by cisternal taps, showed that 1 to 5% remained after 24 h, as opposed to the known clearance of interferon from the bloodstream to this level within minutes. The distributions of interferon and
131
I-labeled albumin were similar as demonstrated by kinetics of clearance monitored over 24 h. Beginning with and after experimental infection of rabbits, daily intraventricular injections of one million units of interferon were given for as long as 3 weeks. Interferon was prepared from cell culture fluids after pressure dialysis and chromatography on Sephadex G-100. This intensive treatment did not prevent encephalitis, but prolonged the length of the incubation period by one- to two-thirds. The outcome after intraventricular administration was not as favorable as when one million units equally divided between intramuscular and intravenous injections were given at the time of challenge. Interferon administered in the subarachnoid space in this fashion is apparently inadequate to protect the rabbit against rabies. Its role as an adjunct measure, or other methods of administration in the nervous system, remains to be examined.
Collagenolytic Activity of Dental Plaque Associated with Periodontal Pathology
Authors: Loesche W. J. , Paunio K. U. , Woolfolk M. P. , Hockett R. N.
Published: January 2020

Certain dental plaques, removed from sites of gingival and periodontal pathology in mentally retarded, institutionalized individuals, when incubated in phosphate buffer with Achilles tendon collagen, gave rise to an increase in ninhydrin-positive material. These plaques, while showing great variability, released significantly more ninhydrin-positive material per milligram of plaque (wet weight) than did either the endogenous or heat-treated controls. Certain plaques could also break down soluble, tritiated, labeled collagen isolated from the calvaria of chicken embryos.
Bacteroides melaninogenicus
and
Clostridia histolyticum
were found in plaques by either fluorescent antibody or cultural methods.
C. histolyticum
, when detected, accounted for about 0.01 to 0.1% of the bacteria in plaque. A conspicuous isolate from some plaques was a
Bacillus
species which rapidly liquefied gelatin. Cell-free supernatants of this organism were able to degrade about 50 to 70% of the soluble collagen when incubated at 36 C.
C. histolyticum
ATCC 8034 caused an 80% degradation of the collagen under the same conditions of incubation. The
Bacillus
strains were facultative, could ferment glucose, reduced nitrate to nitrite, and were catalase, indole, and urease negative. The limited taxonomic information for the isolates is compatible with the description given for
Bacillus cereus.
Mycoplasma-Associated Immunosuppression: Effect on Hemagglutinin Response to Common Antigens in Rabbits
Authors: Bergquist Lois M. , Lau Benjamin H. S. , Winter Charles E.
Published: January 2020

A
Mycoplasma
-associated immunosuppression of the primary and secondary hemagglutinin response to a common gram-negative antigen of
Escherichia coli
(014) was demonstrated in the rabbit when preincubated mixtures of common gram-negative antigen and
Mycoplasma arthritidis
membranes were injected intravenously. A similar immunosuppression was demonstrated only for the secondary hemagglutinin response to a common gram-positive antigen of
Staphylococcus aureus
when preincubated mixtures of common gram-positive antigen and
M. arthritidis
membranes were employed as immunizing materials. The immunosuppressive effect occurred with small quantities of
M. arthritidis
membranes and appeared not to be limited to the host in which arthritogenic properties of the organism were manifested.
Delayed Hypersensitivity Reactions Provoked by Ribosomes from Acid-Fast Bacilli: Physical Characteristics and Immunological Aspects of Core Ribosomal Proteins from
Mycobacterium smegmatis
Authors: Loge R. V. , Hill W. E. , Baker R. E. , Larson C. L.
Published: January 2020

Ribosomal subunits from
Mycobacterium smegmatis
were analyzed by using sedimentation velocity, sedimentation equilibrium, and acrylamide gel electrophoresis experiments. These s
0.w
values for the subunits are 48.7
S
and 28.1
S
. The molecular weight of the 49
S
subunit is about 1.65 × 10
6
, and that of the 28
S
subunit is 7.8 × 10
5
. Both subunits contain about 37% protein and 63% ribonucleic acid. A protein-deficient particle having an s
0.w
value of 15.7
S
contains about 11% protein and 89% ribonucleic acid. Skin tests showed all subunits and proteins to be active as agents in provoking delayed hypersensitivity, but the 16
S
protein-deficient particle, as well as the proteins derived from it, was more specific than the subunits themselves.
Cycloheximide-Resistant Glycosylation in L Cells Infected with
Chlamydia psittaci
Authors: Stokes Gerald V.
Published: January 2020

L cells (mouse fibroblasts), uninfected and infected with the meningopneumonitis strain of
Chlamydia psittaci
, were labeled with [
14
C]glucosamine, and their membranous organelles were separated by isopycnic equilibrium centrifugation of whole cell homogenates on discontinuous sucrose gradients. Glycosylation of host membranes continued throughout the infection. Cycloheximide almost completely inhibited glycosylation in uninfected L cells, but it only partially inhibited the process in infected host cells. Cycloheximide-resistant glycosylation of membrane fractions with [
14
C]glucosamine increased as the infection proceeded and was probably due to the action of chlamydial enzymes. Modification of host membranes by glycosylation may play a role in the natural development of chlamydial infections.