bluuebunny/crossref_metadata_2025 huggingface dataset:
The Purification of Tetanus Toxoid by Ammonium Sulfate Fractionation
Authors: Levine Leo , Stone Joseph L.
Published: January 2023

A simple method for the routine purification of tetanus toxoid for use in prophylactics has been presented. The products have been evaluated with respect to purity, yield, antigenicity, and stability. Of 34 runs, 21 furnished between 40 and 80% of the original activity with a mean increase in purity of 29%. The reported yield and degree of purification of the remaining 13 runs were probably distorted due to non-specific flocculation of the crude toxoids. Useful concentrates of adequate antigenicity and stability were obtained by this method.
Western Equine and St. Louis Encephalitis Viruses in the Blood of Experimentally Infected Wild Birds and Epidemiological Implications of Findings
Authors: Hammon W. McD. , Reeves W. C. , Sather Gladys E.
Published: January 2023

Several species of wild birds from Kern County, California were inoculated subcutaneously with high dilutions of western equine and St. Louis viruses. This frequently resulted in readily detectable viremia beginning within 24 hours after inoculation. With the western equine virus, viremia attained titers up to 10−6 and lasted 3 to 4 days. Antibody developed within 15 to 29 days following inoculation in all those adequately tested. Birds inoculated with St. Louis virus developed much lower titers of viremia, usually detectable only in the undiluted and the 10−1 dilution and only once detected at a level of 10−4. These titers are comparable to those found previously in domestic fowls, and mosquitoes had become infected in repeated instances following feedings made on these fowl. The inoculated birds in these experiments did not always develop neutralizing antibodies during the interval which had proved adequate for the western equine virus. One bird with demonstrated viremia had failed to develop detectable antibodies 30 days after inoculation. It is concluded from these experiments and other observations that in Kern County, California wild birds are as important as and probably more important than domestic fowl as sources of virus infection for mosquitoes.
The Effect of Proflavine on the Growth of Viruses in the Chick Embryo
Authors: Briody B. A. , Stannard Claudia
Published: January 2023

Proflavine, when injected a few hours before or after the inoculation of virus, inhibits the multiplication of Type B strains of influenza virus in embryonated eggs, but does not influence the growth of Type A strains. The drug interferes also with the multiplication of vaccinia virus on the chorioallantois of the chick embryo when injected 2 hours following the virus. The growth of mumps and Newcastle disease viruses is unaffected by concentrations of proflavine active against vaccinia virus and Type B strains of influenza virus. In concentrations found to inhibit viral growth, proflavine is not virucidal, and it does not inhibit hemagglutination by any of these viruses.
Antibody Response of Human Subjects to Epidemic Typhus Vaccine Three to Eight Years after Previous Immunization
Authors: Murray Edward S. , Ofstrock Avis , Snyder John C.
Published: January 2023

Several million Americans have received two or more injections of Cox-type epidemic typhus vaccine during the past nine years (1). This is in accord with the policies of the Armed Forces and various civilian agencies of the United States which require all personnel being sent to potential zones of epidemic typhus to be immunized against this disease. Today, many of these previously immunized individuals are being sent overseas again either as members of the Armed Forces or as civilians participating in the programs of the State Department, the Economic Cooperation Administration, or other agencies.

The question now arises as to whether these previously immunized individuals should receive the usual primary course of two injections of typhus vaccine or whether a single booster dose will suffice to recall and raise their typhus antibodies to the maximum obtainable level (2). Data bearing upon this problem are presented below.2
Studies on Acute Disseminated Encephalomyelitis Produced Experimentally in Rhesus Monkeys
Authors: Kabat Elvin A. , Wolf Abner , Bezer Ada E.
Published: January 2023

Large doses of cortisone prevented the local granulomatous response to the tubercle bacilli and the development of acute disseminated encephalomyelitis in rhesus monkeys injected with brain emulsions with adjuvants.
Studies of a “Complete” and an “Incomplete” Rh Antiserum with cDe/cde and -D-/-D- Cells
Authors: Sturgeon Phillip , Stettner Constance E.
Published: January 2023

Summary and Conclusions

Two samples of a serum obtained from a voluntarily immunized Rh negative male donor during the early phases of his immunization were studied with cDe/cde cells. Titrations of the first sample gave results characteristic of a complete (agglutinating, bivalent) serum. The second sample obtained 70 days later gave results characteristic of an incomplete (univalent) serum. Parallel tests of these two samples with -D-/-D- cells resulted in the second sample's showing, for the most part, the characteristics of a complete serum. This unique property of -D-/-D- cells was described originally by Race, Sangers and Selwyn.

However, the titrations in saline of the second sample with -D-/-D- cells revealed a suggestive prozone and the saline endpoint was considerably lower than the albumin endpoint. These latter observations suggest that the incomplete antibodies in this sample conform to the accepted reaction pattern of univalent antibodies and not bivalent antibodies, even with respect to -D-/-D- cells.

Samples of another high-titer incomplete serum also obtained from a volunteer donor were studied with the above cells. No agglutination in saline occurred with cDe/cde cells; however, a narrow zone of weak agglutination was present with -D-/-D- cells. The incomplete endpoints, however, were 64 times as high. These latter observations, though demonstrating the ability of -D-/-D- cells to be agglutinated by incomplete anti-D sera, again conforms to the accepted reaction pattern of univalent antibodies.

Further studies were carried out with various mixtures of a complete and an incomplete Rh antiserum. The results of these studies strongly suggest that the ability of -D-/-D- cells to be agglutinated in saline by incomplete anti-D sera depended upon the presence in the incomplete antiserum of relatively small amounts of complete (bivalent) antibodies.
The Interaction of Erythrocytes and Antibodies with Synthetic Polymers and Some other Substances
Authors: Frazer Winifred T. , Calvin Melvin
Published: January 2023

The ability of ten polymers and eleven simpler chemical substances to inhibit agglutination of type O Rh0 red cells by corresponding antibodies and type A cells with anti-A serum has been tested in concentrations ranging up to as high as 40 mg/ml. No substance of practical importance was among them.
The Homogeneity of Purified Diphtheria Toxins and Toxoids As Investigated by the Semisolid Precipitin Technic
Authors: Bowen Harry E.
Published: January 2023

Summary and Conclusions

All of 6 purified diphtheria toxins (and toxoids) and 17 antitoxins tested by the semisolid precipitin method demonstrated multiple lines of reaction, the number of lines formed being dependent upon both the antitoxin and toxin used. Of the 3 types of antitoxin investigated, low-salt antitoxin generally gave the largest number of lines with any particular toxin, followed by high-salt and enzyme digested material in that order. This difference is thought to be due to the method of preparation of the various types of antitoxins rather than to any inherent difference between the various fractions. Antitoxins resulting from short-term immunization tend to show fewer lines with any given toxin or toxoid. The number of lines formed by a particular toxin or toxoid with antitoxin appears to be inversely related to the estimated purity of that toxin or toxoid. By appropriate absorption techniques, toxin or toxoid can be deprived of its major line of reaction with a particular antitoxin, without markedly altering its pattern of minor lines in the semisolid precipitin reaction. The above conclusions strongly support Pope's contention that “purified” diphtheria toxins and toxoids may contain numerous antigens other than toxin and toxoid. A crude method of estimating the purity of toxoid preparations by use of the semisolid precipitin reaction is described, and data are presented to demonstrate that values so obtained agree reasonably well with values obtained by Pappenheimer's methods.
Studies on Certain Viruses Isolated in the Tropics of Africa and South America. Immunological Reactions As Determined by Cross-Neutralization Tests
Authors: Smithburn K. C.
Published: January 2023

Seventeen strains of viruses isolated in Africa and South America, all of them neurotropic for Swiss mice, were submitted to immunological investigation by means of cross-neutralization tests, along with a group of 12 previously known viruses having similar neurotropic properties. The results show that: Eleven of the viruses are distinct from each other and from all the agents of the comparison group. These 11 appear, therefore, to be hitherto unknown. They include: Bwamba fever, West Nile, Semliki Forest, Ntaya, Bunyamwera, Uganda S, Zika, Anopheles A, Anopheles B, Wyeomyia and Ilhéus viruses.The Kumba virus, isolated in West Africa, is identical with the Semliki Forest virus, previously isolated in Uganda.Of 5 viruses originally encountered in the Passos area in Brazil, one, designated Heterogeneous virus, has apparently been lost, and the remaining 4 appear to be members of the group (GDVII and FA types) of mouse encephalomyelitis viruses. These are the strains designated Haemagogus A, Sabethes, Leucocelaenus and Haemagogus B.Unilateral or nonreciprocal cross-reactions may occur between viruses which are not identical. Limited investigations of such unilateral reactions are reported and discussed.

Previous investigations showing a close relationship between Mengo and encephalomyocarditis viruses and between Russian spring-summer encephalitis and louping ill viruses are confirmed.

The importance of the hitherto unknown viruses is discussed.
Toxic Manifestations in Rabbits and Mice Associated with the Virus of Western Equine Encephalomyelitis
Authors: Fastier Lyle B.
Published: January 2023

Animals inoculated with high concentrations of the Western equine encephalomyelitis virus exhibited well-defined toxic manifestations directly referable to the virus particle. The toxin resembled those of other toxigenic viruses in producing a febrile reaction and associated leucopenia after the intravenous inoculation of rabbits. Mice inoculated by the same route died within two days with gross necrotic lesions in the liver. There was no evidence that significant viral multiplication was associated with these latter changes. The toxin, relatively stable at 4 C, was destroyed at 56 C for 30 minutes (a procedure simultaneously effecting a ten-thousandfold reduction in the intracerebral infectivity titre). Neutralization of virus infectivity by specific antiserum also abolished clinical evidence of toxicity.

The above results have been incorporated in a discussion on the relationship between viral toxicity and the normal infective process of the same virus.
Q Fever Studies
Authors: Lennette Edwin H. , Clark William H. , Jensen Florence W. , Toomb Carolyn J.
Published: January 2023

Discussion and Summary

The time of appearance, increase in titer, and persistence of complement-fixing antibody against Coxiella burnetii was studied in 102 individuals. In addition, in 85 of the 102 patients, similar observations were made on the agglutinating antibody.

Agglutinating antibody was detectable in the serum of 92% of the patients by the end of the second week, whereas complement-fixing antibody was detectable in only 65 per cent of the cases at this time. By the end of the third week, agglutinating and complement-fixing antibodies were detectable in the same proportion of patients, i.e., 95 and 97% respectively. At this time the median titers of both antibodies were in the range of 1:128–1:256. These findings suggest that the rickettsial agglutination test may have some advantages in the early diagnosis of Q fever. From the economic standpoint, however, the complement-fixation test is more readily adaptable to laboratory use. Also, it has been suggested in other rickettsial diseases that the complement-fixation test may be a better tool for epidemiologic surveys, since complement-fixing antibody has been observed to persist longer than the agglutinating antibody (16). From certain of the data presented here (charts E and F in Figure 3) concerning individual patients, it might also be inferred that in Q fever the complement-fixing antibody persists longer than the agglutinating antibody. However, if the patients are considered as a group (Figures 1 and 2) it would appear that the two antibodies in man are similar in their patterns of persistence; this would suggest that either the agglutination or complement-fixation test may be employed in serologic surveys. Parenthetically, it may be stated that in experimentally-inoculated large animals (sheep), the small amount of data available suggest, in general, that the agglutinating antibody appears early and tends to disappear within a few weeks (17); consequently, in surveys whose purpose is to detect remote as well as recent infections in livestock, it would appear that the complement-fixation test is the preferable method.

The long persistence in man of both the agglutinating and the complement-fixing antibody emphatically underlines the need for caution in the interpretation of the results of serologic diagnostic tests performed on a single specimen of serum. The median titers of both antibodies in our series of patients remained at appreciably high levels for many months, so that a titer obtained on a single serum specimen cannot, in the absence of strong supporting epidemiologic and clinical evidence, be interpreted unequivocally as evidence of current infection with C. burnetii.
The Effect of Chemical Substances in Mice Infected with St. Louis Encephalitis, PR8 Strain of Influenza, or the Lansing Strain of Poliomyelitis Virus
Authors: Wooley Jerald G. , Bond Howard W. , Perrine Theodore D.
Published: January 2023

Studies with T2 Escherichia coli bacteriophage showed that this bacterial virus could be restricted in its multiplication by several types of chemicals (1–2). These studies were pursued further by investigating the effect of many of the chemicals that inhibited the bacteriophage multiplication, analogues of chemicals that reversed the inhibitions, chemicals known to have metabolic activity and their analogues, as well as chemicals of miscellaneous structure for their effect in protecting animals infected with animal viruses. The viruses used in animals consisted of St. Louis encephalitis, PR8 strain of influenza and Lansing strain of poliomyelitis.

Technique. Swiss albino male mice from a pure strain raised at the National Institutes of Health by promiscuous breeding were used. Mice weighing 16 to 18 g each were used. They were housed in glass battery jars, 5 to a jar, and were fed Purina dog chow and water. They were divided into groups of 20 to 40 mice each.
Studies on Herpes Simple Virus
Authors: Jawetz Ernest , Coleman Virginia R.
Published: January 2023

An egg-adapted strain of herpes simplex virus was used in a study of the neutralizing activity of human sera in the embryonated egg. The neutralization test employing lethal endpoints after the injection of virus-serum mixtures into the yolk sac was found to be simpler and more reliable than that employing the reduction of pock count on the chorioallantoic membrane. Undiluted serum neutralized disproportionately more virus than diluted serum. The neutralizing capacity of dilute human sera was greatly enhanced by incubating virus-serum mixtures for 1 hour at 37 C prior to injection. The procedure increased the neutralization index of different sera in varying degree and thus permitted their distinction.
Studies on the Chemical Inhibition of T2r+ Bacteriophage
Authors: Bourke Anne R. , Robbins Mary Louise , Smith Paul K.
Published: January 2023

Twenty-eight out of 180 compounds tested prevented lysis of Escherichia coli B by T2r+ bacteriophage. Possible mechanisms of their activity are presented. Inhibition by many of these compounds was reversed by one or more naturally occurring substances, particularly those associated with oxidative enzyme systems. Quinacrine was antagonized competitively by riboflavin and nicotinic acid, and non-competitively by thiamin. Phthalic acid was reversed non-competitively by thiamin, cysteine and creatinine. Inhibition induced by streptomycin was reversed competitively by pyridoxine and non-competitively by p-aminobenzoic acid.
Quantitative Aspects of Influenza Virus Multiplication
Authors: de St. Groth S. Fazekas , Cairns H. J. F.
Published: January 2023

It is not the subject matter of these studies which unites them in a series but the intent of showing that the present knowledge of influenza virus multiplication lends itself to quantitative and theoretical treatment. The three preceding papers were concerned with different facets of the problem; here an attempt will be made to define and estimate the constants common to them all.

Relation between infectivity and haemagglutinating power. The infectivity of influenza virus preparations is usually expressed as the number of 50%-infective doses (ID50) contained in a standard volume. To find out how many infective units constitute a haemagglutinating dose, we collected the data from the protocols kept by five members of the Hall Institute. Only results fulfilling the following requirements were considered: (a) the test material was grown from dilute seed (b) for less than 48 hours, and (c) both infectivity and haemagglutinin titrations were done on the day of harvest.
Interaction of Protein Antigens and Antibodies
Authors: Turner E. W. , Boyer P. D.
Published: January 2023

Experiments have been conducted on the solubilization and dissociation of specific diphtheria toxoid-antitoxin precipitates by acid and alkaline solutions and by alkaline solutions containing urea and thiocyanate. Additions of urea and thiocyanate were observed to have synergistic effects in solubilizing specific precipitates.

Diphtheria toxoid-antitoxin precipitates could readily be dissolved under conditions which were sufficiently mild to prevent loss of serological activity of both components exposed separately, for example, in 0.1 M glycine buffer, pH 11.0, 2 C. Electrophoretic analyses of such dissolved precipitates showed that they migrated essentially as single components. More drastic conditions resulted in slight separation of the precipitates as measured by electrophoretic analyses. However, diphtheria toxoid and antitoxin exposed separately to these more drastic conditions showed some denaturation as demonstrated by decreased precipitate formation or complexing. These experiments support the suggestion that the forces involved in the interaction of protein antigens and antibodies are similar in nature and magnitude to those responsible for maintaining the native structure of the proteins.

Diffusion experiments with a diphtheria toxoid-antitoxin precipitate dissolved at 2 C in 0.1 M glycine buffer, pH 10.8, containing 1 M urea and 0.1 M KCNS showed that the dissolved precipitate exists under these conditions as large, undissociated aggregates. This result is most readily explained by the theory that antibodies are bivalent.

The results reported together with other data are interpreted as in harmony with the view that successive specific interactions between antigens and antibodies are accompanied by a decrease in affinity so that interactions between aggregates to give precipitates are weaker than the primary interactions between antigens and antibodies.
An Immunologic Study of The Iron-Binding Protein
Authors: Jager B. V. , Gubler C. J.
Published: January 2023

Antisera prepared against two samples of crystalline iron-binding protein were examined as regards homogeneity by means of supernate analyses for simultaneous presence of antigen and antibody, by means of the Oudin diffusion method, and by their characteristics as determined with the quantitative precipitin technique.

Antisera thought to be homogeneous on the basis of these criteria were used for estimation of iron-binding protein in whole human serum. Certain antisera gave values for iron-binding protein that were in good agreement with chemical estimations of this protein. Other antisera, however, gave immunologic values which were in excess of chemical values.

Only by direct comparison of immunological values with chemical values was it possible to select suitable antisera for the immunologic estimation of iron-binding protein.
Effect of Glucose on the Growth of Influenza Virus in Deëmbryonated Eggs and Tissue Cultures
Authors: Daniels Joan B. , Eaton Monroe D. , Perry Maude E.
Published: January 2023

When deprived of glucose, chick embryo tissues infected with influenza virus produced relatively small increments in hemagglutinin and infective virus. This effect of glucose deprivation could be reversed by the addition of glucose to deëmbryonated eggs at 48 hours and to tissue cultures as late as 5 days after infection. In tissue cultures deprived of glucose, oxygen consumption was 60 to 70% of the controls with glucose and deterioration of epithelium was observed in explants after depletion periods longer than four days. Bernkopf's observations on production from concentrated virus inocula of hemagglutinin in the absence of an increase in infective virus were confirmed in deëmbryonated eggs and in tissue cultures. Tissue cultures have been modified, without the use of complex enrichment, to produce virus yields equivalent to those obtained in the deëmbryonated egg.
The Influence of Chemicals on the Propagation of Poliomyelitis Virus in Tissue Culture
Authors: Brown Gordon C. , Schieble Jack H.
Published: January 2023

Various compounds have been tested for their ability to inhibit the propagation of poliomyelitis virus in tissue culture.

Five of nine analogues of amino acids, cysteic acid, dl-ethionine, N-dichloroacetyl-α-(p-nitrophenyl) glycine, α-amino-β-phenylethane sulfonic acid, and β-2-thienylalanine were found to inhibit viral growth.

Benzimidazole, 2,6-diaminopurine, and proflavin, all compounds involved in nucleic acid synthesis, were also effective inhibitors.

The vitamin analogues as a group were ineffective, but one of these, dl-homobiotin, was found to be active.

Glycerol monoacetin, an antagonist of fluoroacetate and phagopedin sigma, an antibiotic, were also found to be inhibitory.

The specificity of action of two analogues of amino acids, ethionine and β-2-thienylalanine, and of two metabolic analogues of purines, benzimidazole and 2,6-diaminopurine, was shown by the partial reversal of their inhibitory effect with the appropriate metabolites. These metabolites, d-methionine, dlphenylalanine, adenine, guanine, and uracil are therefore shown to be essential for the growth of poliomyelitis virus in tissue culture.
Immunochemical Studies with the β-Galactosidase and Structurally Related Proteins of Escherichia Coli
Authors: Cohn Melvin , Torriani Anna Maria
Published: January 2023

Rabbit antisera against purified Escherichia coli β-galactosidase (Gz) have been prepared. These antisera precipitate the enzyme but do not inactivate it. The fact that the enzyme titrates with this antiserum to the same endpoint whether it is determined with lactose or orthonitrophenol β-galactoside as substrate, demonstrates that both activities are associated with a single antigenic species. Inactive extracts (from cells which have not been induced to produce enzyme) as well as active extracts (from cells which possess the enzyme) contain a protein (Pz), inactive as a β-galactosidase which cross-reacts strongly with the antibody to Gz. Pz, although immunologically closely related to Gz is distinguishable by the following properties. Pz will precipitate 80–90% of the antibody directed against Gz, but leave a fraction which will react only with Gz to the exclusion of Pz. In mixtures of Pz and Gz, Gz precipitates preferentially and Pz remains in solution as long as the precipitation of Gz is not complete. Pz is nonantigenic for rabbits whereas Gz is an exceptionally efficient antigen. The following summarizes some other properties of Pz and Gz. Pz and Gz have very similar solubility properties but are separable by the fact that Gz and Pz have appreciable differences in their temperature coefficients of solubility. Gz is more stable than Pz towards the action of heat, ultraviolet irradiation and proteolytic digestion. Gz can be converted into an enzymatically inactive, immunologically identical protein by the action of dilute formalin under proper conditions. These results show that the induced formation (enzymatic adaptation) of β-galactosidase involves the appearance of a new antigen Gz and, therefore, the biosynthesis of a new protein. The significance of the immunological behavior of Pz and Gz is discussed with special reference to problems of protein homogeneity and interpretation of the reactions of complex systems.
Four New Provisional Serotypes of Shigella
Authors: Ewing W. H. , Hucks M. C.
Published: January 2023

Four serotypes which have the biochemical characteristics of typical shigellae are described. It is believed that these serotypes should be considered for possible future addition to the Shigella schema, but for the present should remain sub judice. Cultures of one of the serotypes did not produce acid from mannitol and hence should be considered for addition to subgroup A (Shigella dysenteriae subgroup) of the schema. Cultures of the other three serotypes ferment mannitol, but were not related serologically to members of the Shigella flexneri group and therefore should be considered for addition to the Shigella boydii subgroup (subgroup C). The serologic relationships of the new serotypes to known shigellae and to Escherichia coli O groups are described.
The Quantitative Determination of the Extent of Despeciation of Modified Equine Antitoxin
Authors: Levine Leo , Wyman Louise , Chen Bei-Loo , Murphy Julia
Published: January 2023

A method is presented for estimating the relative reactivity with respect to horse serum of antitoxins processed from horse plasma, by observing skin reactions of sensitized rabbits. It is feasible to test 4 preparations, each on 3 rabbits, with standard fiducial limits of from 53% to 188% of the true value. As the sensitizing dose was increased, the relative reactivity of a given test material appeared greater. The function was asymptotic at higher dose levels. The greater the sensitizing dose, the smaller was the variance of the relative reactivity within its replicates. Movement of control and test reaction diameters in opposite directions of magnitude upon a second skin challenge demonstrated simultaneous partial desensitization to normal horse serum and sensitization to the modified antigen consequent upon the first challenge. Incubation periods between sensitization and skin challenge of 2, 3, and 4 weeks made no difference in values obtained for relative reactivity. Relative skin reactivity of various preparations as determined in sensitized rabbits showed correlation with anaphylactic tests in guinea pigs. The less reactive of 2 materials compared in human subjects was also the less reactive when compared by the rabbit skin test.
Studies on the Cultivation of Poliomyelitis Viruses in Tissue Culture
Authors: Weller Thomas H. , Enders John F. , Robbins Frederick C. , Stoddard Marguerite B.
Published: January 2023

In 1949, in preliminary notes, we recorded the fact that the in vitro propagation of the poliomyelitis viruses may be accomplished in suspended cell cultures of various types of human tissues. The present paper details the procedures we have employed in the preparation and maintenance of suspended cell cultures for this purpose, and summarizes observations on the behavior of poliomyelitis viruses in such cultures, as observed during the past 4 years.

Each of 3 antigenic types of poliomyelitis virus has been propagated in suspended cell cultures of human embryonic skin and muscle tissue, in cultures of human embryonic brain tissue and in cultures of mature human kidney tissue. In addition it has been demonstrated that one or more of these 3 types of virus will multiply in cultures of various other human embryonic tissues, including intestine, adrenal and spleen. Likewise, mature human tissues including thyroid, foreskin, and testis have been successfully employed for the cultivation of the Lansing strain as well as materials from two tumors, an embryoma of the kidney, and a neuroblastoma. Poliomyelitis virus has also been cultivated in monkey kidney cultures.

Assay of the infectivity of materials removed from the tissue cultures was perfomred by the classical method of titration in animals and also by the technique of titration in vitro employing the cytopathogenic effect of the poliomyelitis viruses as an indicator of viral activity. Materials from cultures prepared with human embryonic skin-muscle tissues, kidney tissue, or brain tissues yielded the highest infectivity titers. Serial propagation in suspended cell cultures of human embryonic skin-muscle tissue was achieved for 23 passages with the Lansing strain, for a cumulate period of 331 days in cultivation, and for 15 passages with the Brunhilde strain, during a total period of 267 days. Throughout these prolonged periods of cultivation, virus production in each instance remained essentially constant as assayed by the in vitro technique. The titers as determined in vitro were in most cases equivalent to those observed in the central nervous system of the infected animal. However, in experiments with Lansing virus prolonged cultivation in human tissues was associated with a marked decline in apparent infectivity titer of the agent for the mouse. Likewise, in one experiment with Brunhilde virus, a significant decrease in the apparent infectivity titer for the monkey was observed.
Relationship of Various Antiorgan Sera
Authors: Fleisher Moyer S.
Published: January 2023

In our previous studies (1, 2) concerning tissue specificity we were constantly faced with the fact that the various tissues showed a very great complexity of relationship. In many cases this complexity tended to cloak the evidence of the tissue specificity and it was only by means of comparative, quantitative studies that we were able to prove certain facts. It was, however, evident that tissue specificity could actually be demonstrated in the case of the tissues which were used (namely, liver, kidney, spleen, brain, muscle and testicle). Furthermore, there was noted a relationship between the various organs of animals of the same species, which was evidently due to a species specificity. In addition there were suggestions of a further relationship between the various organs which could not be explained by the simple species specificity.
Studies on the Toxicity of Human Blood Plasma for Guinea-Pigs
Authors: Levinson S. A.
Published: January 2023

I. Introduction. In our previous report (1), we have briefly mentioned the phenomena produced by the intravenous injection of human plasma into the guinea-pig which we associated with coagulation changes. It is the object of this paper to report in some detail the results of this observation, and to show briefly the relationship it has with anaphylaxis and the associated phenomena.

The theories of anaphylaxis and anaphylactic shock are so numerous that it is only possible to mention a few of them, especially those which may have any bearing on our findings. Biedl and Kraus (2), Schultz and Jordan (3) among others have shown that the peculiar and rapid fatal character of anaphylactic shock in the guinea-pig depends on the valve-like closure of the bronchioles, due partly to the special susceptibility of the bronchiolar musculature, and partly to the thick lining of these tubes in the guinea-pig, so that when contraction of the bronchi take place, the lumen is obstructed.
Cutaneous Hypersensitivity in Brucellosis
Authors: Benedict Albert A. , Elberg Sanford S.
Published: January 2023

The use of the surface-active agents, Duponol WA and Lorol, for the preparation of skin test antigens from Brucella suis is described. These reagents were more effective than Tergitol 7, Tween 80, Triton A-20, phenol and water. The time of onset of cutaneous hypersensitivity in guinea pigs was related to the number of living organisms used for sensitization. The intensity of the allergic response reached a maximum after 8 weeks of infection and decreased to a relatively constant level from the tenth to eighteenth week of infection. Reactions of the purpura type were produced in guinea pigs by the crude extract from the fourth to tenth week of infection. Such reactions were not observed in response to the partially purified reagent. The crude extract exhibited specificity for Brucella infection in guinea pigs. Responses in animals infected with Mycobacterium tuberculosis and Coccidioides immitis were not of the same order of magnitude in size or intensity as were produced in experimental Brucella infections. The partial purification and crystallization of the antigen by precipitation at pH 4–5 and ammonium sulfate precipitation at 6–10% of saturation at pH 7.0 and 5 C was described. The antigen is water-soluble near pH 7.0, thermostable and inactivated by trypsin. Tests for the presence of desoxyribonucleic and ribonucleic acids were negative while the tests for protein were positive. Only traces of carbohydrate were detected in the purified antigen. Upon further purification skin test activity of the reagent sharply increased, while the concentration of carbohydrate and of substances responsible for strong ultraviolet absorption at 2700 Å decreased. The final product showed a low specific density at this wave length. Repeated intradermal injections of the allergen did not result in the development of either systemic (anaphylactic) or dermal sensitivity in normal guinea pigs. These intradermal injections of the purified antigen resulted in very minor stimulation of agglutinin formation in guinea pigs and in a small group of humans. Guinea pigs infected subcutaneously with as few as 9 cells of Br. suis responded to the cutaneous test with the purified antigen 1 week following infection. The antigen was about 225% more potent when used to skin-test rabbits infected for 4 weeks with Br. suis (homologous species) than for rabbits infected for 4 weeks with Br. abortus or Br. melitensis (heterologous species). An allergen capable of detecting skin sensitivity in guinea pigs infected with Br. suis could not be extracted from Br. abortus strain A19 under the conditions of the method used successfully on Br. suis. Upon storage at 5 C for 11 months a solution of the purified antigen lost 85% of its skin test potency. Response to dosage data on various preparations were susceptible to analysis of relative potency by virtue of the parallelism of the log dose-log response relationship. The skin-test antigen inhibits the linear migration of leukocytes of normal guinea pigs in direct proportion to its concentration.
A Serologically-Active Erythrocyte-Sensitizing Substance from Typhus Rickettsiae
Authors: Chang Shih-Man
Published: January 2023

A serologically active substance capable of specific sensitization of erythrocytes, the ESS, has been isolated from typhus rickettsiae by boiling typhus rickettsia-infected yolk-sac material in dilute alkali. No such ESS can be isolated from uninfected yolk sac by the same method.
On the Lack of Agreement of the Constant Toxin and Constant Antitoxin Flocculation Reactions of Diphtheria Toxin and Equine Antitoxin
Authors: Bowen Harry E. , Wyman Louise
Published: January 2023

Summary and Conclusions

Six equine antitoxins and 4 toxins and toxoids were investigated, with respect to their flocculating properties. The earlier, graphical presentation of Boyd demonstrating the existence of distinct optima for constant toxin and constant antitoxin procedures was fully confirmed in all cases. The flocculation reaction was equated with respect to total concentration and ratio AT/T with the following general conclusions: Log time of flocculation bears an inverse linear relationship to log concentration of toxin plus antitoxin. Log time of flocculation bears a third-order parabolic relationship to ratio AT/T. The overall speed of reaction and the magnitude of relationships a. and b. are dependent upon both the toxin and antitoxin in question. Evidence is offered to indicate that toxin-antitoxin pairs showing different optima by routine flocculation procedures can be made to give identical optima in constant toxin and constant antitoxin titrations by using procedures which eliminate all pertinent variables except the ratio AT/T. On the basis of these findings the authors suggest that failure to obtain identical results by the constant toxin and constant antitoxin methods is primarily because of failure to control the total concentration of reagents and, of lesser importance, to establish comparable AT/T intervals. For routine purposes, either the constant toxin or the constant antitoxin method can be made most nearly ideal (in the sense of measuring the single variable, AT/T ratio) by using a toxin of about 3.5 times the titer of the antitoxin and ignoring any difference in volume from tube to tube. A more exact procedure would depend upon knowing the specifically precipitable N of the antitoxins and applying this information to adjusting the total concentration (of antitoxin plus toxin) in each tube to a constant value. The latter would be most readily accomplished by diluting the antitoxin to contain the same concentration of specifically precipitable N as the toxin against which it is to be titrated and to proceed with the titration without regard for the final total volume.
Studies on Immunization Against Plague. V. Multiplication and Persistence of Virulent and Avirulent Pasteurella Pestis in Mice and Guinea Pigs
Authors: Walker D. L. , Foster L. E. , Chen T. H. , Larson A. , Meyer K. F.
Published: January 2023

Some avirulent strains of Pasteurella pestis progressively decrease in numbers in host tissue without indication of multiplication, but viable bacilli can still be found up to the 6th to the 9th days after inoculation. Other strains maintain their numbers well for 5 or 6 days and are then progressively eliminated. Some strains multiply and definitely increase in numbers before being overcome and the tissue sterilized. With the avirulent strains studied, at no time were bacilli cultured from the blood later than 15 minutes after injection.

Those strains which are effective in stimulating development of immunity in mice may be the ones that, even though incapable of extensive multiplication, are rich in soluble envelope antigen or may be the ones that, although less productive of envelope antigen, have the capacity to multiply and to attain numbers adequate to provide necessary quantities of envelope antigen.
The Hemagglutinin of St. Louis Encephalitis Virus
Authors: Chanock Robert M. , Sabin Albert B.
Published: January 2023

When physiologic salt solution was used for the extraction of mouse brains infected with St. Louis encephalitis virus, hemagglutinin appeared at varying times after the suspensions were prepared, zone phenomena, as well as incomplete patterns of agglutination, were frequently encountered, and the activity quickly disappeared on storage at 4 C, even when the large particles were removed by centrifugation at 13,000 rpm for 1 hour. When the pH of 7.2 exhibited by unbuffered saline suspensions of mouse brain was changed to 7.5 to 8.5 with dilute NaOH prior to centrifugation, the zone phenomena and incomplete patterns disappeared. It was, furthermore, found that the stability of the hemagglutinin on storage at 4 C could be maintained as long as the pH of the solution remained at about 7.7 or higher, and the larger tissue particles were removed by centrifugation. Physiologic salt solution, buffered at pH 9.0 with borate-KCl in 0.016 M concentration, was found to be an excellent fluid for the extraction of hemagglutinin from infected mouse brains. By its use, hemagglutinin of high potency was regularly demonstrated in 13,000 rpm supernates obtained immediately after preparation of the suspension. No zone phenomena were encountered and the solution could be stored in an ordinary refrigerator for at least 290 days without loss of titer. Even when this optimum method was used for extraction, no hemagglutinin could be obtained from the brains of mice infected with the Webster No. 3 strain of St. Louis encephalitis which has had a very large, although unknown, number of mouse passages over a period of 18 years.
The Hemagglutinin of St. Louis Encephalitis Virus. III. Properties of Normal Inhibitors and Specific Antibody; Use of Hemagglutination-Inhibition for Diagnosis of Infection
Authors: Chanock Robert M. , Sabin Albert B.
Published: January 2023

To overcome the marked instability of the SLE hemagglutinin at the pH that is optimum for hemagglutination, inhibition tests were performed by mixing hemagglutinin diluted at pH 7.7 and 1 C with serum or tissue suspension adjusted to pH 6.5 at room temperature, and immediately adding the erythrocyte suspension. The normal inhibitor in mouse brain was extracted in larger amounts by hypotonic than by isotonic solutions, and was largely associated with particles sedimentable at 13,000 rpm for 1 hour. The normal inhibitor in brain is not destroyed during the course of infection. Human and rabbit sera contain more normal inhibitor than mouse brain extracts. Normal inhibitor in solution can remove hemagglutinin from erythrocytes and reverse hemagglutination without affecting the receptors on the red cells.

The SLE normal inhibitor is only partially removed from serum by extracting with chloroform in a manner that removes practically all of the inhibitor for the Japanese B hemagglutinin. Precipitation and extraction with an appropriate amount of acetone yields a fraction which is practically devoid of normal inhibitor, but contains all the specific hemagglutination-inhibition antibody. The SLE hemagglutinin combines with specific antibody in different proportions from normal inhibitor. For each fourfold increase in concentration of hemagglutinin, the 50% inhibitory titer dropped twofold for specific antibody and fourfold for normal inhibitor. Antisera for the West Nile and Japanese B virus contained specific antibody for the SLE hemagglutinin but in much lower titer than antisera for the SLE virus. Specific hemagglutination-inhibition antibody in high titer was found in 8 patients with St. Louis encephalitis during the acute phase of the illness. Tests on human beings, without history of encephalitis, revealed that the hemagglutination-inhibition antibody was absent among residents in a nonendemic area, and present in low titer only among those residents of an endemic area who also possessed neutralizing antibody for the SLE virus.
The Efficacy of the Intracutaneous Route of Injection and the Susceptibility of the Hartley Strain of Guinea Pigs in Experimental Allergic Encephalitis
Authors: Lipton Murray M. , Freund Jules
Published: January 2023

Sterile disseminated encephalitis was produced by Rivers and his associates in monkeys by injecting CNS tissue using often repeated injections (1). By means of adjuvants consisting of paraffin oil and killed mycobacteria or Nocardia asteroides, rapid production of encephalomyelitis has become possible. This has been demonstrated in monkeys, guinea pigs, mice, rabbits and dogs (2–7). The substance responsible for the induction of allergic encephalitis has been the subject of extensive investigations (8). In such studies, it is essential to find conditions which afford the production of encephalitis by a single injection and in a relatively short and narrow range of time. It is also advantageous to produce easily recognizable clinical symptoms which can be correlated readily with histological findings. It is desirable that a large percentage of the injected animals should respond.

In many of the studies on the fractions from the central nervous tissue, repeated injections have been used.
Characteristics of Antibiotic-Producing Strains of the Ocular Bacterial Flora
Authors: Halbert Seymour P. , Swick Lois , Sonn Constance
Published: January 2023

The antibiotics produced by a series of 22 ocular Micrococci have proven to be heterogeneous. Nine different antibiotic agents were produced by the 22 strains. These were apparently active by at least 3 distinct and independent metabolic mechanisms. Fourteen of the 22 antibiotics were rather clearly of polypeptide nature, as evidenced by enzyme destruction and other properties. The remaining 8 agents were resistant to enzyme activity, but a few observations suggest that these too, could be peptide structures. The antibacterial spectra of samples of these strains have been examined in some detail. Only gram positive species were affected significantly. Micrococcus strains were inhibited most frequently, while Corynebacterium, Sarcina, and Bacillus cultures were also commonly and highly susceptible. Streptococci, Clostridia and Mycobacteria were moderately susceptible. In general, the spectra were grossly similar for the three groups of antibiotics which acted by differing metabolic mechanisms. A high degree of variability was often shown in the susceptibility of strains of a given species to a given antibiotic producer.
Quantitative Studies on Passive Cutaneous Anaphylaxis in the Guinea Pig and Its Relationship to the Arthus Phenomenon
Authors: Ovary Zoltan , Bier Otto G.
Published: January 2023

The induction of passive cutaneous anaphylaxis in the guinea-pig requires a latent period, which for the egg albumin-rabbit anti-egg albumin system can not be reduced to less than 30 minutes. A period of 3 to 6 hours represents the optimum for local sensitization to 0.01 μg anti-Ea N. The binding of circulating antibody by previously intradermally injected antigen does not affect the intensity of passive cutaneous anaphylactic reactions in the guinea pig. Reactions at virgin sites are only observed if large amounts of antibody are used for local sensitization, indicating that enough antibody has passed into the circulation to cause general cutaneous sensitivity. Neoantergan has a definite inhibitory effect on passive cutaneous anaphylactic reactions induced by small amounts of antibody (0.07 μg AbN), but exerts only a slight effect on the reactions caused by large amounts of antibody (30 μg AbN) such as required for the Arthus reaction. Horse antibody, while able to elicit strong Arthus reactions, is entirely ineffective in promoting passive cutaneous anaphylaxis in the guinea pig. The meaning of these findings is briefly discussed in relation to the fundamentally different mechanisms of the Arthus phenomenon and of passive cutaneous anaphylactic reactions.
The Flocculation Reaction of Rabbit Antibody
Authors: Bowen Harry E. , Wyman Louise
Published: January 2023

Summary and Conclusions

The flocculation reactions of 3 anti-pneumococcal polysaccharide and 2 antihuman albumin rabbit sera have been equated with respect to total concentration of reagents, Ab/Ag ratio, and flocculation time, with the following conclusions: Ab/Ag ratio shows a parabolic relationship to log flocculation time. log total concentration of antigen plus antibody is inversely related to log flocculation time. The differences between α and β optima could be resolved, under these experimental conditions, by properly adjusting the concentration in all tubes to a constant. The flocculation optimum obtained by the constant concentration procedure with each antigen-antibody pair examined corresponded to a point in the region of antibody excess. The differences between the flocculation behavior of equine and of rabbit antibody are discussed.