bluuebunny/crossref_metadata_2025 huggingface dataset:
VITAMIN B12Co60 READILY PASSES THE PLACENTA INTO FETAL ORGANS AND NURSING PROVIDES B12 FROM MOTHER TO PUP
Authors: Woods W. D. , Hawkins W. B. , Whipple G. H.
Published: June 2004

The placenta is permeable to B12Co60. Together with salts, sugar, amino acids, vitamins, and proteins, the B12 is contributed to the growing embryos from the mother's body stores. At birth, the placenta contains a liberal amount of B12, which the mother regains by ingestion of the placentas.

Nursing draws a liberal amount of B12Co60 from the mother's stores and contributes it to the body of the pups where it is absorbed and distributed in the various organs, much as noted when adult dogs are injected with B12Co60. A redistribution is noted as the months pass; but the high values in the heart, liver, and gastric mucosa persist, and the brain usually shows a slow increase.

Enterohepatic circulation of B12 may involve bile and enteric content plus hepatic, gastric, and pancreatic epithelial secretion. This type of body recycling of radioactive B12 is discussed but not proven.

High values of B12 in the heart, brain, gastric mucosa, and liver indicate that the vitamin is functionally active, not an inert fraction.
STUDIES ON THE PATHOGENESIS OF RABIES IN INSECTIVOROUS BATS
Authors: Sulkin S. Edward , Allen Rae , Sims Ruth , Krutzsch Philip H. , Kim Chansoo
Published: June 2004

Studies on the influence of environmental temperature on the pathogenesis of rabies in two species of experimentally infected Chiroptera, the Mexican free-tailed bat (Tadarida mexicana) and the little brown bat (Myotis lucifugus), provided evidence that little or no viral multiplication occurs in the inactive host during experimentally induced hibernation. When inoculated animals are wakened from hibernation by transfer to a warm room, virus previously in "cold storage" multiplies, reaching detectable levels in various tissues. Similar results were obtained with two strains of rabies virus, a canine rabies street virus which produced a fatal infection in man and a strain isolated from the pooled brown fat of naturally infected little brown bats. However, certain differences in the characteristics of these virus strains were observed. The canine rabies virus strain produced an encephalitic disease in mice and overt symptoms in bats; the bat rabies virus producing an encephalomyelitic disease in mice and infrequent symptoms in bats. The bat rabies virus had a greater predilection for brown adipose tissue than the canine strain. Results obtained with the bat rabies virus in hibernating animals indicate that after a period of latency in a dormant animal activated virus may reach the salivary gland more rapidly, with greater frequency, and attain higher concentrations than in animals which have not experienced a period of hibernation. The significance of these results as they relate to the natural history of bat rabies is discussed.
EFFECTS OF CHRONIC EXCESS SALT FEEDING
Authors: Dahl L. K.
Published: June 2004

Among female rats and dogs which were fed excess salt for periods of about 1 and 4 years, respectively, elevations in plasma cholesterol concentrations were frequent and in some cases, quite marked.

Rats on a similar regimen for only 2 and 6 months had less frequent and significantly lesser elevations of cholesterol.

The lipid changes did not appear to be related primarily to increased blood pressure, kidney disease, differences in body weight or food consumption.

There is no present evidence which suggests that this phenomenon is related to atherogenesis although it was speculated that this was possible.
FACTORS AFFECTING THE CHAIN LENGTH OF GROUP A STREPTOCOCCI
Authors: Ekstedt Richard D. , Stollerman Gene H.
Published: June 2004

Group A streptococci which grew in long chains in the presence of homologous anti-M antibody were split into their original length by the addition of an excess of homologous M protein to the culture. The chain-splitting reaction showed temperature and pH optima (37°C., 7.5) and was completely inhibited at 0°C. or by heat-killing the long chains at 56°C. prior to the addition of M protein. Addition of sublethal doses of HgCl2, or of penicillin, inhibited the chain-splitting reaction. Pneumococci behaved in entirely comparable fashion to streptococci in similar experiments.

Virulent strains of streptococci formed the shortest chains when broth media was enriched with serum. The chain-shortening effect of serum enrichment of the media was most apparent with encapsulated strains and under cultural conditions that favored capsule formation. Loss of capsules by mutation or by unfavorable growth conditions resulted in increase in chain length. The activity of the chain-splitting mechanism seemed to be independent of M protein, however, since encapsulated M-negative variants also formed very short chain in serum-enriched media. The physical presence of the capsule was not essential for chain shortening since enzymatic removal of the capsule with hyaluronidase during growth did not affect chain length.

These results strongly suggest that chain-splitting of streptococci and pneumococci occurs by an active metabolic mechanism, presumably enzymatic, which is inhibited by the union of surface antigens with specific antibody.
RELATION OF PARTICLE SIZE OF C POLYSACCHARIDE COMPLEXES OF GROUP A STREPTOCOCCI TO TOXIC EFFECTS ON CONNECTIVE TISSUE
Authors: Roberson Bob S. , Schwab John H. , Cromartie William J.
Published: June 2004

The component of Group A streptococci which is responsible for the chronic, remittent, multinodular lesion of connective tissue is derived from the cell wall. Further evidence is given to support the essential role of the group-specific C polysaccharide in the production of this lesion.

A series of particles containing the group-specific C polysaccharide was prepared, ranging in size from large cell wall fragments to the relatively small hapten. A comparison of the lesion producing capacity of the particles in this spectrum revealed that maximum toxic activity is associated with C polysaccharide complexes of intermediate size. The discussion considers colloidal properties associated with C polysaccharide complexes of a certain size, and the influence particle size has on persistence in tissue, as possible explanations of the relationship between the size of the C polysaccharide complex and its ability to produce the chronic lesion of connective tissue.
ON THE ABSORPTION OF BACTERIAL ENDOTOXIN FROM THE GASTRO-INTESTINAL TRACT OF THE NORMAL AND SHOCKED ANIMAL
Authors: Ravin H. A. , Rowley D. , Jenkins C. , Fine J.
Published: June 2004

Coliform-free rabbits fed P32 labeled E. coli 0111B4 prior to the induction of experimental hemorrhagic shock were shown to have a substantial amount of the type-specific 0111B4 antigen in the circulating blood, liver, and spleen, whereas normal rabbits fed the same amount of these bacteria and held under identical conditions, but not exposed to shock, have the antigen within the liver, and occasionally in the kidney, but not in the blood.

That the antigen recovered from the blood and tissues was derived from this specific strain of bacteria was demonstrated by the use of the hemagglutination inhibition reaction, by the absence of cross-reacting antigens in appropriate control animals, and by agreement in the amount of antigen as estimated by two different technics.

Transport of bacterial endotoxin across the intestinal membrane appears to be achieved primarily by passive diffusion.

The accumulation of biologically active endotoxin in the blood and tissues of the shocked animal appears to be due to a reduction in the detoxifying potential of the reticulo-endothelial system, and not to a greater than normal absorption of endotoxin from the intestine.

The absence of toxicity in the specific antigen extracted from normal liver demonstrates that the degradation of endotoxic potency can be achieved without altering the chemical integrity of the polysaccharide moiety of the molecule.

The implications of the hypothesis that there is a continuous but fluctuating absorption of bacterial endotoxin from the intestine are briefly discussed, and the contribution of free circulating bacterial endotoxin of intestinal origin to the fate of the shocked animal is noted.
EVIDENCE FOR A LETHAL ENDOTOXEMIA AS THE FUNDAMENTAL FEATURE OF IRREVERSIBILITY IN THREE TYPES OF TRAUMATIC SHOCK
Authors: Schweinburg F. B. , Fine J.
Published: June 2004

The data here reported (Table I) show that a toxin is present in the blood of animals with two types of irreversible hypovolemic shock. These data also show that although blood volume therapy does not correct the hypovolemia because of continuing loss of plasma at the site of injury, the major factor in the progressive decline and death is the endotoxemia rather than the hypovolemia. This is also true of severe and prolonged hemorrhagic shock that is irreversible to transfusion. The data also show that even when there is bacterial activity at the site of injury, the pool of endotoxin in the intestine is the chief source of the circulating endotoxin.

In all three types of shock, the endotoxemia develops because persisting hypovolemic shock renders the RE system unable to destroy the endotoxin.

The demonstration of an endotoxemia as the cause of irreversibility and death in three types of traumatic shock caused by three different agents suggests that a single pathophysiological mechanism accounts for the phenomenon of irreversibility in all types of traumatic shock.
IN VITRO DETOXIFICATION OF BACTERIAL ENDOTOXIN BY MACROPHAGES
Authors: Rutenburg S. H. , Schweinburg F. B. , Fine J.
Published: June 2004

In the in vitro model described normal serum or plasma alone does not detoxify endotoxin, as determined by the response of the thorotrast-primed rabbit to the injection of the reaction mixture after 1 hour of incubation at 37°C. Partial detoxification, however, is achieved by serum or plasma after 3 hours of incubation of the same reaction mixture, as determined by the chick embryo test.

Macrophages in serum or plasma do detoxify endotoxin, as determined by both tests. Granulocytes, which are also able to detoxify endotoxin in the presence of serum, are less potent than macrophages.

Immune serum alone is much more potent than normal serum alone. Macrophages in immune serum are more potent than immune serum alone.

The relevance of the anti-endotoxic properties of these in vitro systems to the processes in the living animal are in doubt, and must await the results of comparable observations in the living animal.
ENTEROVIRAL RIBONUCLEIC ACID
Authors: Holland John J. , McLaren Leroy C. , Hoyer Bill H. , Syverton Jerome T.
Published: June 2004

Chromatographic behavior of whole type 1 poliovirus and phenol-extracted viral RNA on diethylaminoethyl cellulose columns, as revealed by assay of plaque-forming capacity, indicated that infectious RNA had surface properties markedly different from those of the intact virus.

Infectious RNA of type 1 poliovirus and Coxsackie B1 virus was relatively resistant to heat inactivation as compared to intact virus. Kinetics of inactivation at elevated temperatures were multi-hit in character.

The structure of infectious enterovirus RNA was investigated by treatment with chemical inactivating agents. Urea and guanidine as hydrogen bond-disrupting agents, and mercaptoethanol and thioglycolate as disulfide bond-disrupting agents, and combinations of these did not destroy RNA infectivity whereas hydrogen bond-disrupting treatment inactivated intact virus rapidly. RNA infectivity was not reduced by chloroform extraction alone, or by octanol extraction alone, but was reduced by chloroform-octanol extraction which failed to depolymerize RNA to an extent detectable by ultracentrifugal analysis. Infectivity of type 1 poliovirus and Coxsackie B1 virus RNA was destroyed in accordance with first order kinetics by very dilute solutions of pancreatic ribonuclease, and by purified snake venom phosphodiesterase, but not at all by bacterial alkaline phosphatase. Inactivation by venom diesterase was not accelerated by prior treatment of RNA with bacterial alkaline phosphatase. These results indicated that infectivity of enteroviral RNA resided in a single stranded structure, that a single break of a phosphodiester bond anywhere along the structure was sufficient to destroy infectivity, and that infectivity did not require a terminal phosphate group.

Hydroxylamine, but not other carbonyl reagents, rapidly destroyed infectivity of intact type 1 poliovirus viral RNA without depolymerization of RNA-detectable by behavior in the analytical ultracentrifuge.

With S35-methionine-labeled poliovirus a very small fraction of radioactivity remained in RNA preparations following phenol extraction.

No evidence could be obtained to indicate that infectious enteroviral RNA was composed of subunits.

RNA extracted with phenol during the course of infection of HeLa cells with type 1 poliovirus resembled RNA obtained from purified whole virus with respect to heat inactivation, hydroxylamine inactivation, chromatographic separation, susceptibility to protein denaturing agents, and ability to infect productively both naturally susceptible HeLa cells and naturally insusceptible L strain mouse cells. Intracellular production of infectious RNA paralleled intracellular maturation of whole virus and preceded it by a very short interval.
STUDIES OF TWO KINDS OF VIRUS PARTICLES WHICH COMPRISE INFLUENZA A2 VIRUS STRAINS
Authors: Choppin Purnell W. , Tamm Igor
Published: June 2004

Two kinds of virus particles have been found in varying proportions in influenza A2 strains isolated during the 1957 pandemic. Pure populations of the different particles were obtained, and these substrains were genetically stable on serial passage in the chick embryo.

The two virus particles differ markedly in several biological properties though they are antigenically similar. One kind of particle, designated "+," is relatively sensitive to specific antibody, is highly sensitive to inhibition by serum inhibitors and urinary mucoprotein, fails to elute or elutes very slowly from human erythrocytes, and is capable of agglutinating erythrocytes treated extensively with V. cholerae filtrate. The other particle, designated "-," is relatively insensitive to antibodies and urinary mucoprotein, completely insensitive to serum inhibitors, elutes rapidly from erythrocytes, and can agglutinate erythrocytes treated extensively with V. cholerae filtrate. Both "+" and "-" particles destroy virus receptors on urinary mucoprotein.

The relative proportions of these two particles determine the characteristics of parent strains in reactions with specific antibody, mucoprotein inhibitors, and erythrocytes. The "+" and "-" particles with several easily identifiable markers are well suited for genetic studies.
ON THE RELATION OF THE SIZE OF THE INTRAINTESTINAL POOL OF ENDOTOXIN TO THE DEVELOPMENT OF IRREVERSIBILITY IN HEMORRHAGIC SHOCK
Authors: Wiznitzer T. , Schweinburg F. B. , Atkins N. , Fine J.
Published: June 2004

Additional evidence is presented affirming the role of the intestinal pool of endotoxin in producing irreversibility in prolonged hemorrhagic shock. The fact that coliform-free rabbits tolerate exposure to a degree and duration of hemorrhagic shock which is lethal for rabbits that possess the normal flora, and that these tolerant rabbits lose their tolerance when E. coli are introduced into the gut several hours before inducing shock, demonstrate the critical importance of the size of the pool of endotoxin. That there is a proportionality between the size of the pool of endotoxin and the tolerance of hemorrhagic shock is suggested by the survival rate of several series of coliform-free rabbits fed E. coli by gavage. The rate was less the more firmly the E. coli were reestablished in the flora. The presence of the usual number of coliform bacteria in the intestinal flora does not mean the presence of the usual amount of endotoxin in these bacteria. The amount of endotoxin depends not only on the size of the population, but also, as our own experience demonstrates, on the particular ecological factors extant at any particular time which govern the amount of endotoxin elaborated by any given strain or strains of coliform bacteria.
THE REMOVAL OF CARTILAGE MATRIX BY PAPAIN
Authors: Potter J. L. , McCluskey R. T. , Weissmann G. , Thomas L.
Published: June 2004

In rabbits, the depletion of cartilage matrix which occurs following intravenous administration of papain treated with iodacetamide is attributable to a portion of the enzyme in the disulfide form which has not undergone alkylation. It is this portion that is reactivated in cartilage in vivo and initiates the enzymatic breakdown of the protein-polysaccharide complex which forms a major component of the matrix. Evidence presented in support of these conclusions indicates that, contrary to an earlier hypothesis, papain acetamide is not reactivated in vivo.

Following intravenous injection in amounts up to 4 mg./kg., active and inactive papain leaves the circulation at a rate proportional to the concentration, and it is likely that the initial rate of disappearance represents equilibration with the extracellular space. Following injection in the active or inactive form, a high proportion of papain in serum is bound to protein in the alpha globulin fraction. It is believed that in the case of fully active papain, the proportion which is not bound to alpha globulin becomes attached to other proteins of serum in extracellular fluid, such as albumin, by a process of enzyme substrate combination, and is thus prevented from diffusing into cartilage. In the case of inactive papain, a comparable excess remains free to enter cartilage, where it initiates depletion of matrix following reactivation within the tissue.

These conclusions provide an explanation for the failure of fully active papain to cause depletion of cartilage matrix in vivo; the widespread changes seen after the injection of papain inactivated by iodoacetamide or by simple reversible oxidation are attributable to a small proportion of the injected material which enters cartilage in the disulfide form in a concentration of approximately 2 micrograms/gm. wet weight. The possibility that such a small amount of protease, when reactivated, can produce changes in cartilage matrix has been confirmed by studies on the effects of papain on isolated cartilage and chondromucoprotein in vitro.

It has been shown that severe local injury results when active papain is injected into the skin of a rabbit in low concentrations. Since a much higher concentration of papain can be attained in the circulation without obvious adverse effects, it is evident that binding of the protease by alpha globulin and possibly other serum proteins may exemplify a mechanism whereby the tissues are protected from injury following entry into the circulation of other potentially harmful agents, such as proteolytic enzymes derived from cells or bacteria.
THE PREPARATION AND SOME PROPERTIES OF PURIFIED ANTIBODY SPECIFIC FOR THE 2,4-DINITROPHENYL GROUP
Authors: Farah Fuad S. , Kern Milton , Eisen Herman N.
Published: June 2004

Injection of rabbits with Freund's complete adjuvant containing bovine γ-globulin substituted to near saturation with dinitrophenyl groups leads to the production of large amounts of antibodies specific for the 2,4-dinitrophenyl substituent. These antibodies can be isolated from sera in satisfactory yield (40 per cent) and in a high state of purity (90 per cent specifically precipitable). Hapten dissolution, which was used to screen haptens for their potential usefulness as specific reagents in the purification procedure, provides a means for evaluating, in relative terms, antibody-hapten affinities (in competition with antigen).
THE ISOLATION AND PROPERTIES OF THE SPECIFIC CYTOPLASMIC GRANULES OF RABBIT POLYMORPHONUCLEAR LEUCOCYTES
Authors: Cohn Zanvil A. , Hirsch James G.
Published: June 2004

A method has been described for isolation of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. Homogeneous suspensions of leucocytes were disrupted by lysis in 0.34 M sucrose. This procedure liberated the cytoplasmic contents of the cell and dissolved a considerable proportion of the nuclei. Following disruption, the sucrose lysate was separated into three fractions by differential centrifugation, i.e. 400 g or nuclear pellet, 8,200 g or granule pellet and the postgranule supernate. Microscopic examination revealed that the 8,200 g pellet was composed of intact granules as well as occasional mitochondria. The other two fractions were morphologically heterogeneous.

Studies with isolated granules demonstrated their lysis by a variety of weak acids and surface-active agents. When buffered solutions were employed between the ranges of pH 2.0 and 9.0, granule lysis began at pH 5.5 and was complete at pH 4.0.

Chemical analysis disclosed that the granule pellet contained protein and phospholipid with only traces of nucleic acids.

Approximately 70 to 80 per cent of the total cellular antimicrobial agent phagocytin was present in the granule fraction. This material was liberated from the granules by acid (pH 5.0 or lower).

Studies on selected enzymes showed that acid phosphatase, alkaline phosphatase, nucleotidase, ribonuclease, deoxyribonuclease, and beta glucuronidase were predominantly localized in the granule fraction. Approximately 50 per cent of total cellular lysozyme and cathepsin were also present in the 8,200 g pellet. Disruption of the granules was associated with the release of the majority of granule protein and enzymes in a non-sedimentable form. The properties and composition of rabbit polymorphonuclear leucocyte granules seem to be analogous to those of liver lysosomes.
IMMUNOLOGIC STUDIES OF HEART TISSUE
Authors: Kaplan Melvin H. , Dallenbach Frederick D.
Published: June 2004

Using fluorescent antibody methods, deposits of bound gamma globulin, as determined in unfixed washed sections of auricular appendages from rheumatic hearts, were noted in a significant number (18 per cent) of 100 specimens studied. Such deposits were observed in myofibers, sarcolemma, interstitial connective tissue, and vessel walls. Albumin and fibrin were generally found absent from these sites. Control hearts from normal and pathologic material, including postmortem and biopsied specimens, in general, did not reveal such deposits. These various tissue sites which contained bound gamma globulin frequently exhibited evidence of alteration as indicated both by enhanced affinity for eosin and by strongly positive reaction with the periodic acid-Schiff reagent, and appeared comparable in some cases to "fibrinoid." Bound gamma globulin was not observed in cellular or stromal components of Aschoff lesions, nor was the occurrence of Aschoff lesions correlated with presence of bound gamma globulin. It is suggested that deposition of gamma globulin and the eosinophilic alteration associated with such deposition are related to certain of the pathologic changes of rheumatic heart disease. The nature of such deposits of gamma globulin was considered from immune and non-immune points of view.
THE USE OF PRECIPITIN ANALYSIS IN AGAR FOR THE STUDY OF HUMAN STREPTOCOCCAL INFECTIONS
Authors: Halbert S. P. , Auerbach T.
Published: June 2004

Studies on the purification of group A streptococcal extracellular antigens detectable with naturally occurring human antibodies from normal individuals have been extended. It has been shown that streptococcal electrophoretic fractions intermediate between the most rapidly migrating components are quite complex. In the calcium phosphate chromatography of adjacent electrophoretic fractions, particular antigenic components desorbed at similar buffer elution steps.

It is clear from the results obtained that substantially more extracellular antigens than the twelve heretofore recognized are secreted in human beings during infection, as judged by their detection with human antibodies. The precise number is not yet known, but is probably greater than 16.

Of the nine components which thus far have been separated rather well from the others, four were previously identified as streptolysin "O," diphosphopyridine nucleotidase, proteinase precursor, and desoxyribonuclease B. The accumulated data substantiated these previous identifications.

The identity of a fifth antigen has been made as a possible complex of C carbohydrate and protein. Tentative evidence for the relationship of a sixth component to scarlet fever toxin has been presented.

It has been shown that rechromatography of crystalline proteinase precursor and desoxyribonuclease B on calcium phosphate columns resulted in elution principally at the expected stepwise increase in buffer concentration.

Attempts to isolate antigens present as mixtures in some calcium phosphate chromatographic peaks, by rechromatography on DEAE or CM cellulose columns resulted in only limited further purifications.
IMMUNOLOGIC STUDIES OF HEART TISSUE
Authors: Kaplan Melvin H. , Meyeserian Mary , Kushner Irving
Published: June 2004

Reactions of normal and pathologic sera with heart tissue have been investigated by the immunofluorescent method, with particular reference to presumptive autoantibodies to heart and their differentiation from blood group isoantibodies and Wassermann antibody. In the heart, blood group substances A and B were found distributed in capillary walls, vascular endothelium, and interstitial connective tissue. In surveys of randomly selected sera, isoimmune reactions against tissue blood group substances A and B were noted infrequently. This finding was considered related to the limited sensitivity of fluorescent antibody methods. Heart tissue from blood group O individuals was used for screening of pathologic sera for presence of tissue-reactive factors.

Wassermann antibody was found reactive with constituents of myocardial sarcoplasm, of which the major reactant was cardiolipin. Wassermann-positive sera absorbed with beef cardiolipin gave evidence of reaction with other constituents of myofiber sarcoplasm. Sera of patients with rheumatic fever, rheumatic heart disease, rheumatoid arthritis, disseminated lupus, and liver disease frequently showed a marked reactivity with constituents of myofiber sarcoplasm. These serum factors were differentiated from Wassermann antibody. At least three patterns of immunofluorescent staining could be differentiated by the distribution of reactants in the myofiber sarcoplasm. These reactants were extractable with ethanol and methanol but not by acetone. Sera found reactive by immunofluorescence frequently gave positive flocculation and complement-fixation tests with alcohol extracts of human heart. Immunofluorescent tests were best correlated with flocculation reactions. Sarcoplasmic-reactive factors were associated in some sera with 19S gamma globulin as demonstrated by the use of fluorescent anti-19S gamma globulin. Serologic reactions with homologous or autologous heart were observed particularly frequently with sera from rheumatic patients approximately 2 weeks following cardiac surgery, as well as in some non-rheumatic patients following cardiac or thoracic surgery or acute myocardial infarction. The pathogenetic significance of these presumptive autoantibodies to heart is unknown. As yet, no definite conclusions may be drawn regarding their relationship to bound gamma globulin in rheumatic hearts or to the postcardiotomy and post-infarction syndromes.
STUDIES ON IMMUNE HUMAN HEMOLYSIS
Authors: Hinz Carl F. , Picken Mary E. , Lepow Irwin H.
Published: June 2004

The Donath-Landsteiner reaction was studied using low and high titer antisera and purified antibody, normal and PNH erythrocytes, and human serum complement.

The requirement for complement in both the cold and warm phases of the reaction depended upon the level of antibody used and the sensitivity of the cells to hemolytic antibodies. Complement was not necessary in the cold phase using PNH cells and a potent source of antibody, but complement was required to be present at some stage if hemolysis were to occur.

Optimal conditions for the cold phase were at 1°C. for 30 minutes at pH 7.4. Ca++ ions were required. Hemolysis in the warm phase occurred within one minute, was optimal at 32°C., and required Mg++.

The relation of these observations to previous reports is discussed with respect to discrepant observations on the nature of the Donath-Landsteiner reaction.
STUDIES ON IMMUNE HUMAN HEMOLYSIS
Authors: Hinz Carl F. , Picken Mary E. , Lepow Irwin H.
Published: June 2004

1. The inactivation, of C'1 blocks the completion of the cold phase of the Donath-Landsteiner reaction; inactivation of the other components of complement does not interfere with the cold phase of the reaction.

2. C'2, C'3, and C'4 are required for the completion of the hemolytic reaction. C'4 reacts in either the cold or warm phase, but C'2 and C'3 must react in the warm phase.

3. Partially purified C'1 or C'1 esterase can be substituted for whole serum in the cold phase, although neither reagent contains any of the other components of complement

4. Partially purified serum inhibitor of C'1 esterase blocks the effect of C'1 esterase in the cold phase, and reverses the effect of complement, C'1 or C'1 esterase when incubated with "activated" cells after the cold phase.

5. C'1 esterase activity can be eluted from "activated" erythrocytes with Na3EDTA.

6. The components of human complement in this human hemolytic reaction act in the order C'1, C'4, C'2, C'3. Ca++ is required for the reaction with C'1, and Mg++ is required for the reaction with C'2.

7. Accordingly, a functional role of C'1 esterase in a human disease state is demonstrated, and a human model is established for the study of the mechanism of action of complement.
CHANGING VIRAL SUSCEPTIBILITY OF A HUMAN CELL LINE IN CONTINUOUS CULTIVATION
Authors: Wong Sam C. , Kilbourne Edwin D.
Published: June 2004

During its serial transfer and cultivation in this laboratory, a human conjunctival cell line (Chang) was observed to change in morphology. Concurrently no change was noted in the susceptibility of the cells to viruses capable of infecting the original cell line.

However, it was noted that the derived variant cell line had acquired susceptibility to the induction of cytopathic effects and incomplete virus formation by several strains of influenza viruses.

It was then discovered that swine influenza virus and the N-WS strain of influenza A virus could be serially propagated in the derived cell line with production of infective virus. The swine virus required adaptation, but the N-WS strain did not.

N-WS and swine influenza viruses multiply with infective virus formation only in the variant conjunctival cell and in no other cell line.

Antigenic, cytologic, and virologic evidence is presented that the influenza virus-susceptible variant cell is of human origin and is not a contaminating cell exogenously introduced.

Transition of a cell line from complete insusceptibility to susceptibility to virus infection and multiplication has not been described previously.
STUDIES ON THE PATHOGENESIS OF STAPHYLOCOCCAL INFECTION
Authors: Johnson Joseph E. , Cluff Leighton E. , Goshi Keiichi
Published: June 2004

The influence of repeated staphylococcal infection of rabbit skin upon the characteristics of the experimentally induced lesion was studied. It was found that the repeated infection was associated with the development of delayed hypersensitivity unaccompanied by the appearance of demonstrable serum antibody. The delayed hypersensitivity to the staphylococcus resulted in an increased infectivity of the organism in skin of the sensitized animal, characterized by intensification of the lesions seen with large bacterial inocula and the induction of abscesses with inocula incapable of producing any lesion in normal rabbit skin. Similarly, the severity of experimentally induced pyoarthrosis was greater in sensitized than in normal rabbits. Induction of delayed hypersensitivity by vaccination of rabbits with washed heat-killed staphylococci resulted in the same increased severity of the infection and an increase in infectivity of the microorganism.

In contrast to the observations of cutaneous and joint infection, the sensitized animals appeared to be less susceptible to severe infection of the anterior chamber of the eye.

The role of immunity and hypersensitivity in staphylococcal infection is discussed and the possibility that non-specific inflammation may influence staphylococcal infection in the same way as specific hypersensitivity is indicated. Studies to further elucidate this are presented in the following pages.
STUDIES ON THE PATHOGENESIS OF STAPHYLOCOCCAL INFECTION
Authors: Goshi Keiichi , Cluff Leighton E. , Johnson Joseph E. , Conti C. Richard
Published: June 2004

Studies have been described in which the effect of early and late or established inflammation, upon staphylococcus infection of rabbit skin has been evaluated. Inflammation was produced in skin by thermal, chemical, bacterial and immunological injury, and it was found that the area of inflammation was more susceptible to staphylococcus infection than was normal skin if the bacteria were injected within 2 to 3 days after the injury. When staphylococci were injected into an area of inflammation of over 3 days' duration, there appeared to be an increase in local resistance to infection. The way in which inflammation was produced seemed to have a little influence upon the effects observed. This influence of non-specific inflammation upon staphylococcus infection was compared with the influence of specific bacterial hypersensitivity, which also is associated with an increase in infectivity of the microorganism in sensitized animals. It was concluded that specific bacterial hypersensitivity probably increases susceptibility to infection with the staphylococcus in the same way as non-specific inflammation. The general significance of non-specific inflammation upon infection is also discussed.
FURTHER STUDIES ON THE ROLE OF PROTEASES IN THE ALLERGIC REACTION
Authors: Ungar Georges , Yamura Takuso , Isola Jacqueline B. , Kobrin Sidney
Published: June 2004

Protease activity was measured through the hydrolysis of synthetic amino acid esters in body fluids and tissues of guinea pigs, rats, mice, and humans. Significant in vitro activation was observed in serum and lung slices of sensitized guinea pigs on addition of the specific antigen. Increased proteolytic activity was also seen in reverse anaphylaxis. More marked activation occurred when guinea pig serum was treated with peptone and guinea pig or rat serum was treated with agar. Protease activation was demonstrated in specimens of human skin under the influence of a poison ivy extract or croton oil added in vitro.

Urinary protease activity of guinea pigs increased significantly during the first hours of anaphylactic shock and very markedly in peptone shock. Peptone shock, elicited in mice pretreated with H. pertussis, was accompanied by a considerable increase in protease activity in the peritoneal fluid as compared with non-pretreated mice which were insensitive to peptone.

Proteolytic activity resulting from the activation procedures was due to a number of proteases. The dominant substrate affinity and inhibition patterns suggest that serum and urine proteases are similar to but not identical with plasmin. Anaphylactic activation exhibited patterns different from those resulting from the action of anaphylactoid agents. Tissue enzymes are either of cathepsin- or chymotrypsin-type or mixtures of both.

Some of the activated enzymes, although remarkably effective in hydrolyzing amino acid esters, show no activity on protein substrates. This does not justify, however, their designation as "esterases." They probably belong to the class of specific proteases acting only on a single or a small number of functionally significant protein substrates.

There is at present sufficient evidence to prove not only that protease activation does occur in anaphylaxis and anaphylactoid conditions but also that it is an important component of the chain of reactions leading to the allergic response.
THE MAMMALIAN CELL-VIRUS RELATIONSHIP
Authors: Crowell Richard L. , Syverton Jerome T.
Published: June 2004

Sustained infection of HeLa cells by Coxsackie B3 virus, dependent on presence of viral inhibitor in culture medium, was achieved. Persistent treatment of carrier cultures with anti-Coxsackie B3 hyperimmune monkey serum eventually eliminated virus from carrier cultures indicating that a lysogenic virus-cell relationship was not operative.

Free virus was produced continuously by carrier cultures despite washing and neutralization with antiserum to eliminate free virus temporarily. In carrier cultures, about 1.5 to 1.9 plaque-forming units of virus per cell were cell-associated; approximately 6 per cent of this cell-associated virus was not neutralizable by antiserum. In growth medium containing anti-B3 antibody, cells from carrier cultures formed colonies as efficiently as cells from B3-cured cultures. Assays of carrier cultures for infectious centers indicated that less than 1 per cent of cells produced free infectious virus. The Coxsackie B3 virus-carrier state appeared to represent surface residence of B3 virus on the majority of carrier cells with restriction of productive infection to a small proportion of the population.

Coxsackie B3 carrier HeLa cultures, unlike control cultures, were not destroyed by challenge with Coxsackie B1, B3, or B5 viruses. The B3 carrier state did not interfere with superinfection by herpes, vaccinia, and types 1 to 3 polioviruses. In contrast to parental or B3-cured lines, B3-carrier HeLa cultures superinfected with Coxsackie B1 virus produced no significant virus, and cultures superinfected with B5 viruses produced new virus to a limited extent only.

Specific interference with Coxsackie virus superinfection by the B3-carrier state of HeLa cells was shown to be attributable to failure of attachment in the instance of Coxsackie B1 virus, and failure of penetration and/or eclipse in the instance of B5 virus. The interfering effect was circumvented successfully by superinfection of carrier cells with ribonucleic acid extracted from Coxsackie B1 and B5 viruses.
THE OCCURRENCE OF TWO M ANTIGENS IN CERTAIN GROUP A STREPTOCOCCI RELATED TO TYPE 14
Authors: Wiley Grove G. , Wilson Armine T.
Published: June 2004

A strain of Group A streptococcus previously considered to belong to Type 14 was shown to have two immunologically distinct M antigens, designated Type 14 and Type 51. Most strains having the Type 14 M antigen were found to have also the Type 51 M antigen, and are considered to belong to Type 14–51. Four strains had the Type 51 M antigen without the Type 14 M antigen and one strain had the Type 14 M antigen without the Type 51 M antigen.

The failure of a variant to produce the known M antigen of the parent strain does not necessarily mean that the strain is M-, because a second M antigen may be present as was the case in several strains described here.
ANAPHYLAXIS IN CHOPPED GUINEA PIG LUNG
Authors: Austen K. F. , Brocklehurst W. E.
Published: June 2004

The quantitative release of histamine by specific antigen from perfused, chopped, sensitized guinea pig lung has been used to study the effect of peptidase substrates and inhibitors on the anaphylactic reaction.

The anaphylactic release of histamine is prevented by chymotrypsin substrates and inhibitors but not by trypsin, carboxypeptidase, or leucine aminopeptidase substrates or the soybean trypsin inhibitor. The chymotrypsin substrates and inhibitors appear to be acting on an antigen-antibody-activated step because these substances fail to inhibit if the tissue is washed free of them prior to antigen addition, and because there is complete desensitization of the tissue without histamine release when the antigen is added in the presence of these inhibitors. The inhibitors work equally well in tissue from passively sensitized animals or in tissue from animals actively sensitized with either ovalbumin or bovine gamma globulin. These observations suggest that activation of a chymotrypsin-like enzyme is a necessary condition for the anaphylactic release of histamine in guinea pig lung.

Diisopropylfluophosphate is inhibitory when present at the time of antigen addition but not when the tissue is washed free of unfixed diisopropylfluophosphate prior to adding antigen. This indicates that diisopropylfluophosphate must be acting exclusively on an enzyme which exists in lung tissue in a precursor form resistant to diisopropylfluophosphate until activated by the antigen-antibody interaction.

Thiol alkylating or oxidizing agents also prevent the anaphylactic release of histamine, but in contrast to the situation with diisopropylfluophosphate and the other chymotrypsin inhibitors, the phase of the anaphylactic reaction inhibited by N-ethylmaleimide is available prior to the antigen-antibody interaction.

The similarities and differences between immune hemolysis and anaphylaxis in chopped guinea pig lung are considered in detail.
SPECTRUM AND CHARACTERISTICS OF THE VIRUS INHIBITORY ACTION OF 2-(α-HYDROXYBENZYL)-BENZIMIDAZOLE
Authors: Eggers Hans J. , Tamm Igor
Published: June 2004

2-(α-Hydroxybenzyl)-benzimidazole (HBB) inhibited the cytopathic effects of the following enteroviruses: polio 1 to 3; Coxsackie A9; Coxsackie B 1 to 6; and ECHO virus types 1 to 9, 11 to 21, and 24 to 27. The following enteroviruses were not inhibited: Coxsackie A types 7, 11, 13, 16, and 18; and ECHO types 22, 23, and 28. Other HBB-insusceptible viruses were: arbor B and C, reo 1 to 3; adeno 2 to 4; influenza B; para-influenza 2 and 3; mumps; herpes simplex, and vaccinia.

HBB had no inactivating effect on viral infectivity, but rather inhibited some intracellular step in the reproductive cycle of susceptible viruses. With all viruses examined, inhibition of viral cytopathic effects appeared to be due to inhibition of virus multiplication. Virus inhibition by HBB was demonstrable in monkey kidney, HeLa, and ERK cells.

HBB-susceptible viruses varied quantitatively in their susceptibility to the compound, and different strains of the same virus also exhibited varying susceptibility. No relationship was found between attenuation of polioviruses and their susceptibility to the compound. After passage of HBB-susceptible enteroviruses in the presence of the compound, virus populations with lowered susceptibility to HBB were obtained.

At virus inhibitory concentrations, HBB did not affect the morphology of cells, nor the following cellular metabolic activities: oxygen uptake; glucose utilization; lactic acid production; and incorporation of adenosine into RNA, and of alanine into proteins. The rates of multiplication of HeLa and ERK. cells were not significantly altered by HBB at virus inhibitory concentrations.
THE INACTIVATION OF ENTEROVIRUS INFECTIVITY BY THE SULFHYDRYL REAGENT p-CHLOROMERCURIBENZOATE
Authors: Choppin Purnell W. , Philipson Lennart
Published: June 2004

The infectivity of several enteroviruses was inactivated by the sulfhydryl reagent p-chloromercuribenzoate (PCMB). The rate of inactivation was dependent on the ionic environment in which the reaction was carried out. Inactivation of infectivity was reversed by the thiol compound, reduced glutathione. Under certain conditions, PCMB prevented the adsorption of some enteroviruses to monolayer cultures of monkey kidney cells. The results suggest that enterovirus sulfhydryl groups are involved in the establishment of infection, and that they play a role in the adsorption of virus to host cells.
TRITON HYPERLIPEMIA IN DOGS
Authors: Scanu Angelo , Oriente Pasquale
Published: June 2004

Triton WR-1339, a non-ionic detergent, added to canine serum or to ultracentrifugally separated lipoproteins, induced changes in the lipoproteins which were dependent upon concentration of detergent and class of lipoproteins.

D 1.063 to 1.21 lipoprotein (α-LP) was especially sensitive to the action of triton. Addition of 2 mg. of triton to 1 mg. of α-LP (based on protein content), induced only slight changes in the electrophoretic and flotation characteristics of the lipoprotein. With a tenfold increase of the detergent (triton:α-LP, 20:1), the mixture, analyzed by starch gel and paper electrophoresis, yielded a tritonlipid complex which remained close to the origin, and a nearly lipid-free protein with electrophoretic mobility higher (starch gel) or lower (paper) than native α-LP. The splitting of the lipid and protein moieties of α-LP could not be clearly shown when the same mixture was analyzed by free boundary electrophoresis. Triton alone moved only slightly in an electrical field (paper, starch gel, Tiselius); it sedimented during ultracentrifugation at D 1.006 and D 1.063 and floated at D 1.21.

D 1.006 to 1.063 lipoproteins (ß-LP), required larger amounts of triton to show changes. These were evident in 40 to 80:1 mixtures of triton and ß-LP. In starch gel and paper electrophoresis triton retained, in a position close to the origin, part of the lipids of ß-LP; the remaining ß-LP fraction, impoverished of lipids, had electrophoretic mobility similar to native ß-LP. The triton-lipid complex sedimented at D 1.063.

After addition of triton to complexes [chylomicron-α-P-I131] or [lipomul-α-LP-1131], the electrophoretic and ultracentrifugal analyses of these mixtures revealed that the labeled protein was removed from the triglyceride component. Triton also prevented the occurrence of the interaction between lipomul and α-LP and the hydrolysis of both chylomicrons and lipomul-α-LP by lipoprotein lipase.

It is postulated that, if the changes in lipoproteins and chylomicrons observed in vitro occur in vivo, they could account, at least in part, for the hyperlipemia which develops in animals following administration of triton.
HUMORAL ASPECTS OF THE IMMUNE RESPONSE TO HOMOGRAFTS
Authors: Jensen Emmerik , Stetson Chandler A.
Published: June 2004

Isoimmune sera, prepared in BALB/c mice by immunization with living or lyophilized tissue from C57Bl/6 mice, were found to possess both hemagglutinating and cytotoxic activity for donor cells. Comparative titrations indicated a good correspondence between these two activities. Absorbtion with living or dead donor tissue caused removal or proportionate reduction of both activities. The results are consistent with the existence in isoimmune sera of a single class of isoantibodies, capable of agglutinating donor cells or of sensitizing them for complement lysis. Living and lyophilized tissue differ quantitatively, but not qualitatively, in their capacity to induce formation of these antibodies.
A NEW MOUSE VIRUS CAUSING NECROSIS OF THE THYMUS IN NEWBORN MICE
Authors: Rowe Wallace P. , Capps Worth I.
Published: June 2004

A new mouse virus has been recovered from laboratory and wild mice. The agent induces a non-fatal disease in infant mice, characterized by acute massive necrosis of the thymic medulla and cortex, granulomatous reaction, and subsequent restoration of essentially normal architecture with scarring. Intranuclear inclusion bodies are produced. Virus may persist in tissues of convalescent mice for many months. Electron micrographs of acutely infected thymuses showed nuclear and cytoplasmic karyoannular particles and masses of parallel fibrils in nuclei.