bluuebunny/crossref_metadata_2025 huggingface dataset:
THE INCIDENCE AND PATHOGENESIS OF MYOCARDITIS IN RABBITS AFTER GROUP A STREPTOCOCCAL PHARYNGEAL INFECTIONS
Authors: Glaser Robert J. , Thomas Wilbur A. , Morse Stephen I. , Darnell James E. ,   With the Technical Assistance of Barbara B. Pesch
Published: June 2004

Rabbits subjected to single pharyngeal infections with group A streptococci developed cardiac lesions characterized by myofiber necrosis and a non-granulocytic cellular reaction with histiocytes, lymphocytes, and Anitschkow myocytes. The histopathologic changes were demonstrable in some animals within 24 hours of inoculation, apparently were maximal 72 hours after induction of infection (at which time they were seen in the hearts of all nine rabbits studied), and thereafter healed in the course of the following 2 weeks. The extent of involvement was variable, and with healing the necrotic areas were replaced by fibrous tissue.

When intradermal infections with the same organisms were produced in rabbits, cardiac lesions, indistinguishable from those observed in the pharyngeally infected group, appeared in a much smaller number of animals.

The hearts of five of six rabbits sacrificed a month or more following the last of a series of streptococcal pharyngeal infections exhibited lesions characterized chiefly by fibrosis, although mononuclear cellular infiltrations were also noted. In these repetitively infected animals the presence of occasional multinucleated giant cells and a few small foci of calcification were features not encountered in the single infection group. In a second series of rabbits sacrificed 3 days after the last of three pharyngeal infections with different strains of streptococci, acute as well as more chronic changes were observed.

In none of the lesions in rabbits subjected to single or multiple streptococcal infections were bacteria demonstrable, either in histologic sections or in cultures of myocardial tissue.

A large number of control animals was studied concomitantly, and in only one instance was a lesion, considered comparable to those described in the streptococcal series, encountered.

The implications of these findings, particularly in terms of the non-suppurative sequelae of streptococcal infections in man, are discussed.
CLONAL GROWTH OF MAMMALIAN CELLS IN VITRO
Authors: Puck Theodore T. , Marcus Philip I. , Cieciura Steven J.
Published: June 2004

Two methods for simple and rapid plating of single HeLa cells, human, carcinomatous cells, are described. These result in growth and formation of colonies from each single cell. One of these procedures uses irradiated, non-multiplying "feeder" cells to condition the medium. The second requires more gentle handling of the cells, but otherwise is virtually the same as that used in plating bacteria on semisolid, nutrient media.

By extension of these methods, it is possible to isolate single mutant colonies and grow pure clonal stocks of animal cells. These genetically uniform strains are much more homogeneous in their behavior than the parental HeLa cell population.

Growth curves obtained from developing colonies are highly reproducible. The most active mutant stocks so far isolated display a generation time of 18 to 20 hours.

In pooled human serum HeLa cells assume a highly stretched, ameboid form, with marked motility; whereas growth of the same cells in a variety of non-human sera results in tightly packed, columnar, epithelial-like morphology. The two cell types possess volumes, nuclear cross-sections, plating efficiencies, and generation times which are identical within experimental error, but display widely different cross-sectional areas, suggesting that the basic change occurs in the cell surface. It is conceivable that this change may be related to that which enables the cells of a compact tumor to become invasive.

Animal cells subjected to the standard trypsinization procedures which involve mechanical trauma and repeated washings in incomplete media leak large amounts of P and suffer impaired ability to reproduce as isolated cells.

Application of the methods described in this paper as a tool for quantitative study of normal mammalian cell growth, physiology, genetics, and biochemistry, and the response of cells to drugs, viruses, high energy radiation, and other agents have been indicated.
QUANTITATIVE STUDIES ON THE PROPERDIN-COMPLEMENT SYSTEM
Authors: Leon Myron A. ,   With the Technical Assistance Of Edna Chapman
Published: June 2004

Quantitative methods have been developed for the estimation of C'3 and properdin. With these methods, the kinetics of the over-all reaction between properdin, zymosan, and C'3 were investigated. The reaction followed first order kinetics, provided zymosan was in excess, and the serum not diluted beyond 1/4. The reaction rate was proportional to the temperature, and an activation energy of 20,500 calories/mole was found.

The distinction between total properdin and available serum properdin is discussed.
THE PURIFICATION OF HYPERTENSIN II
Authors: Skeggs Leonard T. , Kahn Joseph R. , Shumway Norman P.
Published: June 2004

The enzymatic conversion of hypertensin I to hypertensin II is described together with the subsequent purification of the product by means of counter-current distribution.

Improved methods are also presented for the preparation of renin and its substrate, as well as in methods for the reaction of these materials and the purification of the resulting hypertensin I.
INFLUENZA VIRUS AND ITS MUCOPROTEIN SUBSTRATE IN THE CHORIOALLANTOIC MEMBRANE OF THE CHICK EMBRYO
Authors: Schlesinger R. Walter , Karr H. Valerie
Published: June 2004

As a preliminary to a study of the fate of mucoprotein substrate in tissues infected with influenza virus, some characteristics of soluble hemagglutination inhibitors (HI) extracted from chorioallantoic membranes (CAM) have been investigated. The inhibitory material was found to be heat-stable, precipitable with cold ethanol, and subject to progressive inactivation by active viruses or by receptor-destroying enzyme (RDE).

Certain changes in the slope of titration curves obtained with precipitated and non-precipitated fractions upon alcohol fractionation suggested that the HI in heated CAM extracts was heterogeneous. Alcohol in low concentration precipitated preferentially the more efficient (longer?) HI leaving the "weak" component in the non-precipitated fraction. With higher concentrations of alcobol, "strong" HI were converted to "weak" ones, either by denaturation or as result of reduced solubility. These changes in slope of titration curves were reflected in significant discrepancies when densitometric titers (HI50) of fractions were compared with their titers in pattern tests.

The action of active viruses on HI, on the other hand, did not induce qualitative changes in the composition of the inhibitory principle: Titration curves retained parallel slopes even when their position was markedly displaced from that of control curves. In its essential properties, the HI from the CAM appeared to be similar to inhibitors isolated from various other biological sources which have been identified as mucoprotein in nature.

Standards for reproducibility of inhibitory titers obtained by the densitometric method of Hirst and Pickels have been presented.

The relation of HI to the allantoic epithelium has been analyzed. It has been concluded that the HI is a normal constituent and secretion product of these cells. Under physiological conditions, i.e. in the intact egg, the HI contained in the mucoid outer layer of allantoic cells appears to be protected from enzymatic action from without, although adsorption of viral particles may be temporarily impeded. In deembryonated eggs, or in excised membranes, a reduction in total inhibitory substrate as a result of the action of RDE has been observed. It has been shown in experiments on adsorption of active or heat-inactivated virus on the allantoic membrane that prevention of adsorption by RDE may require the synergistic action of the active viral enzyme itself.

It has been concluded that the maintenance or restoration of a normal supply of mucoprotein substrate is a function of the ability of allantoic cells to maintain homeostatic conditions under stress.
EFFECT OF SILICATE ON GRAM STAINING AND VIABILITY OF PNEUMOCOCCI AND OTHER BACTERIA
Authors: MacLeod Colin M. , Roe Amy S.
Published: June 2004

Application of silicate solutions to living or heat-killed pneumococci and to certain "viridans" streptococci causes their conversion from a Gram-positive to a Gram-negative state. The original staining properties can be restored by suspending the silicate-treated bacteria in alkaline solutions of various salts but not by simple washing in water.

Living pneumococci and the strains of streptococci whose staining properties are similarly affected are killed when suspended in silicate solutions.

In other Gram-positive species silicate causes conversion to Gram negativity but restoration to positivity occurs upon washing in water. In a third group of Gram-positive organisms silicate has no effect on the Gram reaction. The viability of organisms in these two groups is unaffected by silicate under the conditions employed. No effect on staining or viability of Gram-negative bacteria has been observed.

The effects of silicate on staining and viability are inhibited by nutrient broth or whole serum but not by purified serum albumin. Lecithin, choline, and other substituted ammonium compounds also inhibit the effects of silicate on pneumococci.
AN EXPERIMENTAL ANALYSIS OF THE CURATIVE ACTION OF PENICILLIN IN ACUTE BACTERIAL INFECTIONS
Authors: Smith Mary Ruth , Wood W. Barry
Published: June 2004

Type I pneumococci injected into the leg muscles of otherwise normal mice reached a maximum total population of approximately 106 organisms. In mice rendered severely leucopenic by previous irradiation the maximum bacterial counts recorded were of the order of 109. Since the lesions in the latter animals were relatively acellular, the thousandfold difference in the two experiments represented a rough measure of the antibacterial action of the leucocytic exudate.

The suppressive effect of the leucocytic exudate was shown by histologie studies to involve phagocytosis. The ingestion of pneumococci was clearly demonstrable within the first 12 to 18 hours. Accordingly, it was attributed to surface phagocytosis. In support of this conclusion was the finding that type III pneumococci reached a significantly higher total population in the myositis lesions than did type I. The type III strain used had been previously shown to be resistant to surface phagocytosis during active growth, whereas the type I strain was known to be susceptible throughout its growth phase. Evidence was also presented that the dense leucocytic exudate probably caused in addition a significant degree of bacteriostasis.

When penicillin therapy was begun 9 hours after inoculation, the pneumococci were cleared from the lesions with equal rapidity regardless of the presence or absence of leucocytic exudate. At this early stage the pneumococci were multiplying rapidly in the lesions of both the irradiated and unirradiated mice and therefore were promptly killed by the direct action of the penicillin. When the start of treatment was delayed, however, until 24 hours after inoculation, the bacteria in both sets of lesions had already reached their maximum counts and therefore were presumably resistant to the bactericidal effect of the antibiotic. Under such circumstances the destruction of the bacteria was found to be significantly less prompt in the acellular lesions than in those with a normal cellular exudate.

It is concluded from these findings that, in established pneumococcal myositis in mice, the curative effect of penicillin is due, not to the bactericidal action of the antibiotic alone, but rather to the combined effect of the drug and the cellular defenses of the host. The same conclusion also appears to be applicable to analogous acute infections in man, particularly when they are sufficiently advanced to be definitively diagnosed.
STUDIES ON RESISTANCE TO BACTERIAL INFECTIONS IN ANIMALS INFECTED WITH RICKETTSIAE
Authors: Owen Cora R. , Larson Carl L.
Published: June 2004

Data are presented, demonstrating that infection with Rickettsia typhi brings about a reduction of the death rates and a prolongation of the time of survival in animals subsequently challenged with Pasteurella pestis or Pasteurella tularensis.

This interference with bacterial infection by previous rickettsial infection does not appear immediately after injection of the rickettsiae; it begins to appear around 16 hours after this injection and becomes more marked during the first 96 hours; later it decreases.

The phenomenon is essentially a local tissue reaction with weaker systemic effects.

The phenomenon of interference can be overcome by challenge with too large a number or too virulent a strain of bacteria.

A strain of P. pestis subjected to 10 passages through mice infected with Rickettsia typhi failed to develop resistance to the interfering activity of the latter microbial species. This does not rule out, but might be evidence against, an assumption that the interfering action is due to antibiotic effects from the rickettsiae.
STUDIES ON THE INTERACTION BETWEEN PHAGOCYTES AND TUBERCLE BACILLI
Authors: Stähelin Hartmann , Karnovsky Manfred L. , Suter Emanuel
Published: June 2004

Tubercle bacilli labelled with C14 were prepared by growth on radioactive substrates such as glycerol, CO2, and acetate. These organisms were exposed in vitro to leucocytes (mostly polymorphonuclear leucocytes) from peritoneal exudates of guinea pigs. The respiration of the leucocytes and of the bacilli, alone and together, was followed by determining oxygen uptake and C14O2 production. When heat-killed labelled tubercle bacilli were exposed to leucocytes there was little or no degradation of bacillary material to C14O2 by leucocytic enzymes. On the other hand, conversion of components of sonically disrupted bacilli to C14O2 by leucocytes was significant.

It was possible to determine the oxygen uptake and C14O2 production of phagocytized living tubercle bacilli, and it was found that after phagocytosis the bacilli maintained their rates of oxygen consumption and C14O2 production. This finding was in contrast to observations made with Mycobacterium phlei, a saprophytic acid-fast organism, and with Bacillus subtilis. In these cases oxygen consumption and C14O2 production declined after phagocytosis, and bacterial components were converted to carbon dioxide to a significant degree by leucocytic enzymes.
REVERSIBLE CHANGES IN THE SUSCEPTIBILITY OF MICE TO BACTERIAL INFECTIONS
Authors: Dubos René J. , Schaedler Russell W.
Published: June 2004

Mice were injected intraperitoneally with one of the following bacterial products having endotoxin activity: pertussis vaccine, a suspension of heat-killed cells of Klebsiella pneumoniae (type C), or a purified lipopolysaccharide prepared from cultures of Salmonella typhosa.

Following treatment with either one of these materials, the animals were infected intravenously with virulent cultures of coagulase-positive staphylococci, with bovine tubercle bacilli, or Friedländer bacilli.

The effect of treatment with endotoxin materials on resistance to Friedländer bacilli, staphylococci, or tubercle bacilli was estimated by observing the mortality rates in infected animals, and by determining quantitatively the numbers of living bacteria in the organs at different periods of time after infection.

It was found that mice receiving the infective dose of virulent culture a few hours after treatment with the endotoxin material, were usually more susceptible to infection than were untreated animals. In contrast, mice infected at a later period proved far more resistant to infection than did untreated animals.

The duration of the negative and of the positive phase of resistance was affected by the amount of endotoxin injected. Marked increase in resistance of mice to infection with staphylococci or tubercle bacilli was still evident several weeks after treatment with pertussis vaccine or with purified lipopolysaccharide extracted from typhoid bacilli.
THE AMINO ACID COMPOSITION OF HYPERTENSIN II AND ITS BIOCHEMICAL RELATIONSHIP TO HYPERTENSIN I
Authors: Lentz Kenneth E. , Skeggs Leonard T. , Woods Kenneth R. , Kahn Joseph R. , Shumway Norman P.
Published: June 2004

Preparations of hypertensin II, obtained from the treatment of hypertensin I by the action of the hypertensin converting enzyme of plasma and purified by countercurrent distribution, were quantitatively analyzed for their amino acid content. Chromatography on ion exchange columns showed the presence of equimolar amounts of aspartic acid, proline, valine, isoleucine, tyrosine, phenylalanine, histidine, and arginine.

Hypertensin I was found to contain one mole of leucine and one mole of histidine in addition to the amino acids of hypertensin II. These two amino acids were isolated from the conversion products of hypertensin I and identified as the peptide histidylleucine.

Carboxypeptidase digestion of hypertensin I showed the carboxyl terminal sequence of amino acids to be residue-phenylalanyl-histidylleucine. Similar studies of hypertensin II demonstrated residue-phenylalanine. It was concluded that the conversion of hypertensin I by the plasma hypertensin converting enzyme involved hydrolysis of the phenylalanyl-histidine bond to form hypertensin II and histidylleucine. The further removal by carboxypeptidase of phenylalanine from hypertensin II destroyed all of the vasoconstrictor activity.
CHARACTERIZATION OF INFLUENZA ANTIBODIES BY SERUM ABSORPTION
Authors: Jensen Keith E. , Davenport Fred M. , Hennessy Albert V. , Francis Thomas
Published: June 2004

Absorptions of sera from groups of persons, both normal and after vaccination, resulted in complete removal of antibody to all strains of influenza virus within a type when a strain of antigenic configuration similar to that presumed to be the strain of first experience was employed. Absorption of these sera with strains encountered after the primary antigenic stimulus, removed antibody to strains recently experienced, but failed to absorb the primary antibody. By these methods, three age groups could be distinguished whose characteristic principal antibody was oriented to react with maximal efficiency either with FM1 or PR8 or Swine/1976 influenza viruses. Persons in these age groups had an initial experience with strains of the respective antigenic characteristics found in the variants of influenza virus mentioned. Basis for understanding these results was obtained with sera from successively infected ferrets, when serum absorptions demonstrated all type-specific antibody was oriented to react only with the strain of first infection. In view of this evidence there can now be little doubt as to the marked persistent influence an antigenic experience with influenza virus has upon the antibody-forming mechanisms of the virgin host.
REVERSIBLE COLLAPSE OF RABBIT EARS AFTER INTRAVENOUS PAPAIN, AND PREVENTION OF RECOVERY BY CORTISONE
Authors: Thomas Lewis
Published: June 2004

A substance has been demonstrated in solutions of crude papain, which, when injected intravenously into 1 kilo rabbits, in amounts less than 5 mg., results in complete collapse of both ears. The phenomenon becomes visible 4 hours after injection, and is complete within 24 hours. 3 or 4 days after papain, the ears gradually reassume their normal form.

Ear collapse is associated with depletion of the ear cartilage matrix, and the disappearance of basophilia from the matrix. Similar changes occur in all other cartilage tissues, including bones, joints, larynx, trachea, and bronchi. At the time when the ears are restored to normal shape, the basophilic matrix reappears in cartilage.

Repeated injections of papain, over a period of 2 or 3 weeks, bring about immunity to the phenomenon of ear collapse.

When the arterial circulation to one ear is occluded for 15 minutes at the time of injection of papain, this ear is protected against collapse.

The effect of crude papain could not be reproduced by crystalline papain protease or crystalline papain lysozyme, which together comprise a considerable portion of the dry weight of papain. The nature of the responsible factor has not been determined, and the possibility that chymopapain may be implicated is currently under study.

Cortisone prevents the return of papain-collapsed ears to their normal shape and rigidity. Possibly this reflects a capacity of cortisone to impede the synthesis or deposition of sulfated mucopolysaccharides in tissues.
STUDIES ON TUBERCLE BACILLUS-MONOCYTE RELATIONSHIP
Authors: Fong Jacob , Schneider Patricia , Elberg Sanford S.
Published: June 2004

Studies of the conditions necessary for maintenance of constant cell populations in vitro in the Mackaness type of culture chamber have indicated the importance of preliminary trypsinization of cells and the beneficial effect of 40 per cent rabbit serum in Tyrode's solution. Under these optimal conditions, uninfected suspensions of monocytes exhibited little change in cell numbers over a period of 40 to 72 hours.

Infection of monocytes with the virulent H37Rv strain of tubercle bacillus resulted in an early degeneration of a certain proportion of the cells cultivated in the presence of normal rabbit serum. This degeneration was apparent not only for cells of tuberculin-negative animals but also for those derived from tuberculin-positive animals.

The serum of animals vaccinated with BCG exerted a favorable effect upon the survival of monocytes infected with virulent tubercle bacilli. Treatment with this serum caused a delay in degeneration of infected normal cells (cells of tuberculin-negative rabbits) and a complete inhibition of degeneration of infected immune cells (cells of rabbits vaccinated with BCG).
COMPARATIVE BIOCHEMICAL STUDIES ON NORMAL AND ON POLIOMYELITIS VIRUS-INFECTED TISSUE CULTURES
Authors: Kovacs Ernest
Published: June 2004

Experimental evidence is presented for drastic changes in phosphomonoesterase activities of tissue cultures, brought about by infection with poliomyelitis viruses. Acid phosphatase activity went through a maximum before decreasing almost to zero level. Alkaline phosphatase activity diminished progressively to zero, then with disruption of the cells attamed normal levels. Various aspects of the kinetics were investigated and illustrated. The initial increase of acid phosphatase, in contrast with the alkaline, may mean that the reactions catalyzed by this enzyme continue during the early phase. This period is the time of intense virus production and therefore it was supposed that this enzyme may play some role in virus synthesis. It was assumed that the virus acts as a particle of molecular size and becomes associated with the enzyme complex physicochemically or chemically. This association ends with the disintegration of the host cells. During the cell-virus interaction a toxin may develop which is a strong and general enzyme inhibitor. Various enzyme systems differ in sensitivity toward these virus effects; for instance, acid phosphatase is irreversibly inhibited or may be destroyed. The visible CPE of virus is preceded by a drastic reduction of enzyme activities in whole TC and in its various fractions, which may suggest causal relationship in the mechanism of cell destruction. In arrested or latent infection these processes are operative, but on a smaller scale. The drop in activities cannot be explained by the reduction of tissue mass, which is the consequence, rather than the cause, of enzyme changes. Besides the theoretical significance of these observations the following practical points can be summarized:

1. Changes in phosphatase activities are most strikingly demonstrated in whole tissue cultures inoculated with poliomyelitis virus.

2. There is causal relationship among infection, enzyme changes, and transformation of cell physiology.

3. The biochemical approach provides a quantitative measure of the extent of cell damage, before visible CPE is detectible.

4. Unapparent and active infections with poliomyelitis virus could be differentiated from normal controls by this method.

5. By various manipulations (freezing, long incubation) the difference between normal and infected TC can be enhanced. Suitable technical methods were proposed for various types of investigations.
STUDIES ON THE TRANSFER OF LYMPH NODE CELLS
Authors: Harris Susanna , Harris T. N. , Ogburn Clifton A. , Farber Miriam B.
Published: June 2004

Shigella paradysenteriae organisms were incubated in a solution of trypsin and then removed from the suspension by Seitz filtration. Serologic tests with anti whole Shigella serum indicated that the filtrate contained antigenic material derived from the organisms. When lymph node cells from rabbits not previously injected with Shigella were incubated in vitro with such filtrates and then transferred to irradiated recipients, agglutinins to Shigella appeared in the sera of the latter. The transfer of heated suspensions of cells was not followed by the appearance of agglutinins in irradiated recipient rabbits. Non-irradiated recipients of lymph node cells incubated with the filtrate also developed agglutinin titers, to a lower level than irradiated animals. Agglutinins did not appear in the sera of the majority of non-irradiated recipients of heated cells.

The addition of excess antiserum to either the filtrate or the cell suspension before the incubation of these two materials resulted in a marked reduction in the subsequent agglutinin titers of recipient animals. However, if antiserum was added at the conclusion of the usual 30 minute incubation of cells and filtrate, or even after 5 minutes of incubation, there was no reduction in the agglutinin titers of the recipients.

Cytologic examination of aliquots of a number of the suspensions of cells prepared for transfer revealed that approximately 98 to 99 per cent of the cells belonged to the lymphocytic series.
THE EFFECT OF MALNUTRITION ON THE SUSCEPTIBILITY OF THE HOST TO VIRAL INFECTION
Authors: Sprunt Douglas H. , Flanigan Clyde C.
Published: June 2004

The effect of progressive long term dietary protein depletion on viral susceptibility was investigated in 2 host-virus systems: (1) swine influenza in the male CF1 mouse, and (2) Rous sarcoma virus in the New Hampshire red chicken. Data are presented demonstrating a relationship between host protein nutrition and susceptibility to virus infection. This relationship is shown to be cyclic in character, involving phases of increased and decreased viral susceptibility. The relative resistance of the host on low protein intake is a function of the duration on incomplete diet administration before virus inoculation, and consequently a function of the host's state of depletion. As illustrated in Fig. 6, the cyclic susceptibility change demonstrated by these animals on low protein diet was characterized by an initial phase of increased susceptibility, a secondary phase of increased resistance, and a final phase of increased susceptibility. It is proposed that these alterations in relative viral susceptibility result from metabolic changes occurring within the host during the process of dietary protein depletion. The resistance changes are roughly correlated to periods of depot fat utilization (increased susceptibility), reserve protein utilization (decreased susceptibility), and tissue breakdown subsequent to protein starvation (increased susceptibility).

Many previously published concepts of the interplay of viral susceptibility and host nutrition maintained that host malnourishment led to increased host resistance. The cyclic change in resistance, reported herein, is given as evidence that the effect of host deficiency cannot be explained simply on the basis of an inhibition of virus growth due to retarded cellular metabolism in the host. Protein deficiency is shown not to produce an "all-or-none" effect, but a series of reproducible phases of increased and decreased resistance.

From the aforementioned results it is proposed that the phases of viral susceptibility seen in the protein-deficient host are demonstrative of the dynamic interrelationship between the physiologic state and the resistance of the host. Dietary influences in the normal host, by producing similar metabolic changes, could have analogous implications on innate resistance. It is believed that the foregoing leads to a more clear and dynamic concept of viral resistance in the normal individual.
THE ROLE OF EPINEPHRINE IN THE REACTIONS PRODUCED BY THE ENDOTOXINS OF GRAM-NEGATIVE BACTERIA
Authors: Thomas Lewis
Published: June 2004

Extensive lesions of dermal hemorrhagic necrosis occurred in rabbits when epinephrine (or norepinephrine) was injected into the skin within 4 hours after an intravenous injection of endotoxin. As little as 5 µg. of intradermal epinephrine, and 1 µg. of intravenous endotoxin, were sufficient to produce lesions.

Similar lesions, but smaller in size and surrounded by a zone of acute inflammation, were produced by intradermal injection of a mixture of comparable amounts of endotoxin and epinephrine.

No lesions were produced by combinations of endotoxin with serotonin, pitressin, or ephedrine.

Both types of epinephrine-endotoxin lesion were prevented by pretreatment with cortisone, dibenzyline, and chlorpromazine. They were not prevented by heparin or nitrogen mustard. The lesions produced by intradermal mixtures of epinephrine and endotoxin were greatly enhanced in size and severity in animals treated with nitrogen mustard.

Both types of lesion were prevented in rabbits rendered "tolerant" by repeated injections of sublethal amounts of endotoxin.

It is concluded that endotoxin has the property of altering the reactivity of blood vessels to epinephrine in such a way that this hormone becomes a potent necrotizing agent. The possibility that this effect may represent a basic mechanism in the various intoxicating actions of endotoxin, and certain implications of this hypothesis, are discussed.
THE ENDOTOXIC PROPERTIES OF LYSATES OF GROUP A HEMOLYTIC STREPTOCOCCI
Authors: Stetson Chandler A.
Published: June 2004

Suspensions of group A streptococci were subjected to mechanical disruption by shaking with glass beads. The supernatant solutions produced severe delayed inflammatory reactions on intradermal injection into normal rabbits. Intravenous injections caused systemic reactions characterized by fever, leucopenia, and peripheral vasomotor phenomena. These effects were more pronounced in rabbits which had previously been infected with group A streptococci.

The local and generalized Shwartzman phenomena were reproduced with these crude streptococcal lysates, and the data suggest that streptococci possess an endotoxin similar to those of Gram-negative bacterial species.
EXPERIMENTAL COMBINED VIRAL AND BACTERIAL INFECTION (INFLUENZA C AND HEMOPHILUS INFLUENZAE, TYPE B) IN EMBRYONATED EGGS
Authors: Buddingh G. John
Published: June 2004

Influenza C virus, J.J. strain, is readily propagated following intra-amniotic inoculation of embryonated eggs on the 14th day of incubation. The resulting infection is inapparent in that there is no obvious interference with normal embryonic development and no evidences of injury can be detected by the light microscope.

Hemophilus influenzae, type b thrives luxuriantly in the amniotic fluid of embryonated eggs inoculated by the intra-amniotic route on the 15th day of incubation. The effects of the establishment of the bacterial infection in the embryo are noted by the occurrence of death, bacteriemia or characteristic inflammatory lesions in the form of purulent sinusitis, pharyngitis, tracheo-bronchitis and meningo-encephalitis. These lesions may occur singly or in various combinations. The incidence and severity of disease manifestations in infected embryos depends on the proportion of encapsulated and virulent bacilli in the inoculum, the number of infectious doses and the growth rate of the bacteria in the surrounding amniotic fluid.

Combined viral and bacterial infection established by intra-amniotic inoculation with influenza C virus on the 14th followed by Hemophilus influenzae, type b on the 15th incubation day brings about a significant increase in the incidence and severity of disease manifestations in the embryos. Selective survival and marked acceleration of the growth rate of encapsulated and virulent elements of the bacterial population is promoted in the virus-infected embryos. The disease process appears to be entirely attributable to the bacterial component. There seems to be relatively little or no effect on influenza C virus by the accompanying proliferation of Hemophilus. The exact nature of the virus-induced influences which enhance the pathogenicity of the bacteria and favor the establishment of the infectious process under these circumstances remains to be determined.
STUDIES ON TUBERCLE BACILLUS-MONOCYTE RELATIONSHIP
Authors: Fong Jacob , Schneider Patricia , Elberg Sanford S. ,   With the Technical Assistance Of James Akiyama
Published: June 2004

Studies of virulent, attenuated, and avirulent strains of tubercle bacilli have demonstrated the proficiency of virulent strains to effect degeneration of normal monocytes cultivated in the presence of normal serum. Attenuated strains were less active in this respect, and avirulent bacilli failed to induce monocytic degeneration.

Comparison of the effects of virulent H37Rv with O.T., P.P.D., and a culture filtrate of H37Rv revealed a similarity in action of H37Rv and its filtrate. The action of O.T. and P.P.D. differed from that of H37Rv in that the greatest effect of H37Rv was upon normal cells as opposed to the effect of O.T. and P.P.D. upon immune cells. Additionally, it was demonstrated that immune serum (anti-BCG) protected immune cells against H37Rv but not against O.T. or P.P.D.

The protection of immune cells by heterologous antisera (anti-Salmonella and anti-ovalbumin) as well as by homologous antiserum (anti-BCG) against the degenerative effects of H37Rv indicated a non-specificity in action of serum factors. The ability of the monocytes of animals immunized with BCG and the failure of monocytes of animals immunized with Salmonella rutgers to withstand parasitization with H37Rv, when both types of monocytes were cultivated in immune (anti-BCG) serum, indicated a specificity of cellular resistance.
STUDIES ON BACTERIEMIA
Authors: Rogers David E. , Melly Marian Ann
Published: June 2004

Large numbers of E. coli were rapidly removed from the blood stream of rabbits at rates which initially paralleled the removal of similar numbers of staphylococci. Splanchnic tissues removed approximately two-thirds of the circulating bacilli in transit through the liver and spleen.

In contrast to the cessation of splanchnic trapping noted 20 to 40 minutes following the injection of staphylococci, splanchnic trapping of E. coli continued unchanged for 3 to 5 hours unless the splanchnic tissues were dearly reseeding the blood stream. This resulted in the continued disappearance of E. coli over a 60 to 90 minute period, and differed from the constant bacteriemia maintained beyond 20 minutes after the injection of staphylococci.

Some of the differences in the initial clearance of these two microorganisms appeared to relate to differences in host leukocyte-bacterium relationships. In vitro studies indicated that E. coli were rapidly killed following ingestion by rabbit polymorphonuclear leukocytes. Staphylococci survive such ingestion. The injection of E. coli was followed by a prolonged granulocytopenia with evidence of sequestration of granulocytes within the pulmonary vascular bed. The injection of staphylococci was followed by a transient leukopenia, with rapid return of granulocytes to the circulation (15).

It appears probable that E. coli ingested by sequestered leukocytes are destroyed within the cell, and that leukocytes do not reenter the circulation containing living E. coli. Such intraleukocytic residence in the blood stream has been shown to be of possible importance in the maintenance of staphylococcal bacteriemia (14).

An increasing E. coli bacteriemia occurred rapidly after the initial clearance period, indicating that many sequestered bacilli remained viable. Increasing bacteriemia also occurs 3 to 5 hours after the injection of staphylococci. The bacterial or host cell mechanisms which allow this secondary resurgence of bacterial populations are currently under investigation.
POLYMORPHONUCLEAR INVASION OF WOUNDED CORNEAS
Authors: Weimar Virginia
Published: June 2004

Polymorphonuclear invasion of the wounded rat cornea is quantitatively described. The inflammatory cells enter the wounded tissue during the 5th postoperative hour. They steadily increase in number until they reach a maximum between 24 and 36 hours and return to normal by about the 6th day.

Six hour wounds are used to evaluate the influence of topically applied drugs on the polymorphonuclear infiltration of the wounded area. Sodium salicylate is an effective inhibitor of polymorphonuclear invasion when it is used during the 2nd postoperative hour, but it is without effect when it is applied after this time. Soybean trypsin inhibitor prevents the invasion of polymorphonuclear cells when it is administered during the 2nd hour only, or the 3rd and 4th hours together, or the 5th and 6th hours together.

The results indicate that activation of a proteolytic enzyme (or enzymes) in the injured corneal tissue is an essential step leading to polymorphonuclear invasion.

During the first 2 postoperative hours the anti-inflammatory effects of both drugs are nearly identical and may reflect an inhibition of the same proteolytic reaction or some different but closely related reactions.
EXAMINATION OF THE L FORMS OF GROUP A STREPTOCOCCI FOR THE GROUP-SPECIFIC POLYSACCHARIDE AND M PROTEIN
Authors: Sharp John T. , Hijmans W. , Dienes L.
Published: June 2004

Two strains of L forms of group A streptococci were examined for group-specific polysaccharide and found to lack this substance. One of these was found to make a substance that had several properties in common with M protein. It is suggested that the absence of the cell wall polysaccharide is responsible for the lack of rigidity of the L form and that the L form of this species closely resembles protoplasts as prepared from other species.
IMMUNOHISTOCHEMICAL STUDIES ON THE INTERACTION BETWEEN EHRLICH ASCITES TUMOR CELLS AND NEWCASTLE DISEASE VIRUS
Authors: Prince Alfred M. , Ginsberg Harold S.
Published: June 2004

Newcastle disease virus infection of Ehrlich ascites tumor cells resulted, after a period of time, in the appearance of intracellular viral antigen which could be demonstrated by the fluorescent antibody technique. This antigen appeared in the cytoplasm of infected cells only after inoculation of cell-virus mixtures into the peritoneal cavities of mice. The latent period prior to the appearance of antigen depended inversely on the number of viral particles adsorbed onto the cells prior to inoculation. The final intensity of staining appeared not to be proportionate to the number of viral particles adsorbed to each cell. The appearance of this antigen was not correlated with a rise of titer of infectious, hemagglutinating, or complement-fixing virus. Viral antigen was demonstrated on the surface of tumor cells after adsorption of NDV onto these cells at 0°C. At appropriate virus:cell ratios, antigen was noted to disappear from the surface at 37°C. in vitro, and in vivo, in the absence of demonstrable elution of virus. The appearance of intracellular viral antigen could not be detected in vitro when tumor cell-NDV mixtures were incubated at 37°C., even when an average of 1550 "infectious particles" had adsorbed to each cell.
VITAMIN E DEFICIENCY IN THE MONKEY
Authors: Dinning James S. , Day Paul L.
Published: June 2004

Eight young rhesus monkeys were fed a purified diet devoid of vitamin E. After from 6 to 13 months of feeding, all the animals developed signs of vitamin E deficiency. The signs of vitamin E deficiency in the monkey include muscular dystrophy, elevated excretion of creatine, allantoin, and free amino acids and decreased excretion of creatinine. The vitamin E-deficient monkeys all developed anemia and granulocytosis. Anemia was the first sign of vitamin E deficiency which was observed. All of these signs of vitamin E deficiency were reversed by treatment with α-tocopherol.
DIRECT AND INDIRECT EFFECTS OF X-RADIATION ON ANTIBODY-PRODUCING CELLS
Authors: Dixon Frank J. , Roberts James C. , Weigle William O.
Published: June 2004

X-radiation appears to exert its inhibitory effect on the antibody response by two mutually dependent routes: (a) direct radiation injury to the antibody-producing lymphoid tissue, and (b) indirect effects of altered homeostasis in the radiated host on antibody-producing tissues. Neither of these two effects alone produces significant inhibition of the secondary antibody response made by transferred lymphoid cells. However, 400 to 500 r administered in vitro to the transferred cells, plus 400 r whole body x-radiation of the recipient prior to transfer, completely inhibited the antibody response.
BACTERICIDAL ACTIVITY OF RAT LEUCOCYTIC EXTRACTS
Authors: Fishman M. , Cole L. J. , Silverman M. S.
Published: June 2004

The bactericidal substance present in mitochondrial extracts from rat leucocytes was found to be non-dialyzable, fractionated at 30 per cent (NH4SO4 saturation and was able to withstand temperatures up to 100°C.

The tentative identification of a lipoprotein as the responsible bactericidal substance is based on various observations; an ultraviolet absorption at a peak associated with alpha, beta unsaturated fatty acids, 205 to 208 mµ, inactivation of the bactericidal substance by ether treatment at –70°C. but not at 0°C. and inhibition of the bactericidal activity with lipase treatment as well as with trypsin.
CHEMICAL FORM OF CIRCULATING ANTIDIURETIC HORMONE IN RATS
Authors: Thorn Niels A. , Silver Lawrence
Published: June 2004

Serum, taken from rats under ether anesthesia after injection of hypertonic saline, contained antidiuretic activity in a concentration equivalent to 1 to 3 millipressor units of pitressin per ml.

The active material was retained by a collodion ultrafilter and, on electrophoretic analysis, moved with the beta globulins. Dialysis against 0.2 N acetic acid appeared to dissociate the complex.

The antidiuretic factor was found in serum after stimuli known to cause release of pituitary antidiuretic hormone. Inactivation by thioglycolic acid and by proteolytic enzymes paralleled the inactivation of standard pitressin. Control experiments made it unlikely that serotonin, or any other substance released in the clotting of blood, could have contributed to the antidiuretic action. For these reasons it was concluded that the factor studied was native pituitary antidiuretic hormone circulating as a definite protein-peptide complex.
EXPERIMENTAL URIC ACID NEPHRITIS IN THE RABBIT
Authors: Smith Joseph F. , Lee Yin Chen
Published: June 2004

Experimental uric acid lesions in the kidney of the rabbit have been reinvestigated using the technique of miscrodissection for location of lesions. The maximal initial lesion was found to occur in the first part of the collecting, and not in the ascending Henle and the distal convoluted tubule. Such lesions were only produced by very high dosage (approximately 0.5–1 gm. per kg.).

It was concluded that the argument by analogy with the uric acid lesion cannot be used to support a hypothetical toxic damage to the segments of ascending Henle and the distal convolution in the crush syndrome.
LATENT VIRAL INFECTION OF CELLS IN TISSUE CULTURE
Authors: Morgan Herbert R. , Bader John P.
Published: June 2004

By maintaining L cells in a balanced salt solution of inorganic salts and glucose (BSS) for 2 days or more, they are rendered incapable of supporting the growth of psittacosis virus (6BC), though it infects such cells and is present intracellularly for as long as 3 days in a non-infectious phase. The addition of an enriched medium to such a culture of cells at any time up to 4 days after infection results in the appearance of infectious virus within these cells, which multiplies and is released from the cells, providing the entire period of exposure of such cells to the BSS does not exceed 6 days, following which the cells die. A latent infection with psittacosis virus in a non-infectious phase has been established in a pure line of cells which possess properties of malignancy.
NEUROPATHOGENICITY OF GROUP A COXSACKIE VIRUSES
Authors: Dalldorf Gilbert
Published: June 2004

Coxsackie virus, Group A, Type 14, has been adapted to adult mice and monkeys and induces in them poliomyelitis-like lesions and, in the case of the mice, flaccid paralysis.
THE PATHOGENESIS OF HEMOPHILIA
Authors: Graham John B. , Barrow Emily M.
Published: June 2004

It has been possible to duplicate in the hemophilic dog four of the major experiments which have suggested in humans an "anticephalin" hypothesis for the pathogenesis of hemophilia. The experiments in the dog have been considerably extended, as compared with the human experiments, by a variety of techniques.

I. Asbestos was placed in contact with hemophilic dog plasma, and the clotting time became shorter. When transfused, this plasma had no effect on the defective prothrombin utilization of hemophilic dogs, in contrast to untreated normal plasma.

II. The ionic strength of native dog plasma and dog plasma citrated (38 per cent sodium citrate) then recalcified (0.2 M CaCl2) were calculated. The ionic strength of the native plasma was approximately 0.15 while that of the citrated plasma was approximately 0.21.

Conductivity and freezing point determinations on the plasmas described above were consistent with the idea that the ionic strength of the citrated plasma was significantly higher.

The biphasic dilution curve, to which much significance has been attached in arriving at the "anticephalin" hypothesis, can be produced readily in the dog.

Diluting dog plasma with "iso-ionic" or "hyper-ionic" NaCl solution abolished the biphasic phenomenon. Dilution with distilled water exaggerated the biphasic curve. These experiments suggest that the biphasic curve is an artifact of uncontrolled ionic strength.

III. The prothrombin utilization rates of undiluted whole hemophilic dog blood and hemophilic dog blood diluted 1:2 with 0.85 per cent NaCl were found to be the same.

IV. Ether extraction of both normal and hemophilic dog plasma removed fibrinogen and reduced somewhat the concentration of prothrombin. In treated normal plasma AHF was reduced to the level of untreated hemophilic plasma, thus producing a quasi-hemophilic plasma.

Defibrination and ether extraction of both normal and hemophilic dog plasma "generated" clotting activity which shortened the clotting time of hemophilic plasma and was active in the thromboplastin generation test.

The activity "generated" by defibrination and ether extraction of dog plasma was adsorbed by a BaSO4 suspension and shown, therefore, not to be the anti-hemophilic factor (AHF).

Transfusion of ether-extracted normal or hemophilic dog plasma into hemophilic dogs had no effect on the prothrombin utilization rate, unlike untreated normal plasma which had a marked effect.

Thus, four of the main lines of evidence supporting the "anticephalin" hypothesis were duplicated in the dog. However, by extending the experiments it was found that all were explainable on bases other than the presence of "anticephalin." Such an hypothesis is not necessary, therefore, to explain the pathogenesis of canine hemophilia. The apparent identity of hemophilia in the two species suggests that the hypothesis is also not applicable to humans.