bluuebunny/crossref_metadata_2025 huggingface dataset:
OCCURRENCE OF FOCAL THREE-DIMENSIONAL PROLIFERATION IN CULTURED HUMAN CELLS AFTER PROLONGED INFECTION WITH HERPES SIMPLEX VIRUS
Authors: Hinze Harry C. , Walker Duard L.
Published: June 2004

The occurrence of focal three-dimensional proliferation was observed in human conjunctiva cells in culture after prolonged infection with herpes simplex virus at 30°C. and in KB cells infected with herpes virus at 37°C. for several months in the presence of pooled human serum. This atypical growth pattern consisted of many discrete piles of cells 15 to 20 cells thick scattered over a monolayer of cells.

Three-dimensional growth appeared only in the presence of 10 to 20 per cent of selected human sera. When horse serum was substituted for human serum growth was in an ordinary monolayer.

The abnormal growth pattern of this cell line (C-4 line) has remained stable through continuous culture for over 2 years and was observed consistently in each of 25 clones isolated from the parent culture.

Repeated tests for the presence of persisting infectious virus and for herpes virus antigen in the cells were negative.

Comparison of the C-4 cell line with control conjunctiva cells revealed no significant difference in multiplication rate or in range of viral susceptibility, but the C-4 line was found to be more sensitive to ultraviolet irradiation.
STUDIES ON ANEMIA IN F1 HYBRID MICE INJECTED WITH PARENTAL STRAIN LYMPHOID CELLS
Authors: Harriss Eileen , Currie Cicely , Kriss Joseph P. , Kaplan Henry S.
Published: June 2004

The survival of 51Cr-labeled erythrocytes has been studied in F1 hybrid mice in which wasting disease was produced by injection of parental lymphoid cells taken either from lymph nodes and thymus or from the spleen. Coincident with the development of the disease syndrome, there occurred a severe anemia accompanied by a sudden loss of circulating labeled erythrocytes, whether host or parental. This finding suggests that the anemia is not due solely to specific immunologic reaction of donor tissue against host erythrocytes.
STUDIES ON FLUORESCENT ANTIBODY STAINING
Authors: Goldstein Gerald , Slizys Irene S. , Chase Merrill W.
Published: June 2004

1. A study has been made of the non-specific fluorescent staining of splenic imprints treated with fluorescent sheep antibody globulins.

2. In tissue imprints made with the spleens of antigen-stimulated animals, no morphological distinction was evident between areas showing non-specific fluorescence and specific fluorescence.

3. Elimination of non-specific fluorescence was not achieved by any one, or any combination of the following: (a) conjugating only gamma globulins with fluorescein isothiocyanate; (b) removal of dialyzable fluorescent products on sephadex, followed by concentration through the use of pressure dialysis; (c) use of crystalline preparations of fluorescein isothiocyanate.

4. Individual preparations of fluorescent antibodies were separated by gradient elution chromatography on diethylaminoethyl (DEAE) cellulose into fractions possessing different numbers of fluorescein radicals per molecule of globulin.

5. The coupling ratio of 50 mg fluorescein isothiocyanate (FITC) per gm of protein, as commonly advocated, can not be recommended for the precise localization of antibody globulin in tissues owing to the capacity of the coupled products to give non-specific fluorescent staining. When crystalline preparations of FITC are used instead of the amorphous product at 50 mg/gm protein, far too high non-specific fluorescence results.

6. A fraction with bright specific fluorescence and no or negligible nonspecific fluorescence was obtained from each fluorescent antibody that was prepared by using 6 to 8 mg of crystalline fluorescein isothiocyanate per gm of globulin and was then subjected to DEAE-cellulose chromatography and gradient elution to eliminate the most highly coupled molecules.
CHANGES IN CONNECTIVE TISSUE AND INTESTINE CAUSED BY VITAMIN A IN AMPHIBIA, AND THEIR ACCELERATION BY HYDROCORTISONE
Authors: Weissmann Gerald
Published: June 2004

In view of the theory that an excess of vitamin A causes release of cathepsins from intracellular lysosomes, hypervitaminosis A was induced orally in the larvae of Xenopus laevis. It was predicted that the tails of these amphibia would undergo resorption prior to metamorphosis, since the presence of abundant lysosomes, associated with measurable increases of catheptic activity, had previously been demonstrated in the resorbing tails of amphibia during metamorphosis. This prediction was confirmed; after 3 to 4 weeks of hypervitaminosis A, the tails of treated animals underwent partial resorption.

Other transitory appendages, the rostral tentacles, collapsed after 2 weeks of treatment with an excess of vitamin A, an effect analogous to the collapse of rabbits' ears after intravenous papain. These effects were related to the loss of metachromatic extracellular material in these appendages. Excess of vitamin A caused kyphoscoliosis and prognathos in the larvae.

The hypervitaminotic larvae always developed a mucinous diarrhea, which was associated with a remarkable overgrowth of metachromatic goblet cells of the intestine. The entire intestine of the treated animals was more advanced in development than that of control larvae at equivalent stages.

All the effects of hypervitaminosis A were accelerated by the simultaneous administration of hydrocortisone. This was held to be due to liberation of vitamin A from hepatic stores by the steroid, and is in contrast to the retardation of hypervitaminosis A by hydrocortisone in vitro.
INVOLVEMENT OF ADRENERGIC FACTORS IN THE EFFECTS OF BACTERIAL ENDOTOXIN
Authors: Gourzis James T. , Hollenberg Murray W. , Nickerson Mark
Published: June 2004

The possible involvement of adrenergic mechanisms in the effects of Escherichia coli endotoxin was investigated in several preparations. Appropriate pretreatment of rabbits with E. coli endotoxin significantly increased pressor responses to epinephrine and norepinephrine as compared to untreated controls. Exposure of isolated rabbit aorta strips to E. coli endotoxin in a medium containing whole blood or cellular constituents of blood significantly increased the response to epinephrine. Endotoxin had no effect on responses to epinephrine in ritro when plain Krebs-Ringer solution was used.

Pretreatment with reserpine or phenoxybenzamine (dibenzyline) protected rabbits and mice against the acute lethal effects of E. coli endotoxin. The time period intervening between reserpine or dibenzyline administration and challenge by endotoxin precluded a direct antiendotoxic action of these agents. In addition, incubation of dibenzyline with endotoxin in vitro, under conditions which would favor reaction, did not decrease the toxicity of the latter.

These results indicate that peripheral adrenergic mechanisms are intimately involved in the effects of E. coli endotoxin and support the concept that deleterious effects of endotoxin in shock probably are due to exaggeration of existing vasoconstriction in an already compromised organism.
FURTHER CHARACTERIZATION OF THE ADENOVIRUS ERYTHROCYTE RECEPTOR-MODIFYING FACTOR
Authors: Kasel Julius A. , Rowe Wallace P. , Nemes John L.
Published: June 2004

Agglutinability of human erythrocytes for 3 hemagglutinating adenoviruses was markedly reduced by pretreatment of red cells with a factor present in tissue cultures which had been infected with adenovirus types 1, 2,4, or 15.

The factor responsible for erythrocyte receptor modification was non-dialyzable and unaffected by the action of ribonuclease, desoxyribonuclease, trypsin, chymotrypsin, or ether. The factor was smaller, more thermostable, and separable from the infectious virus.

Erythrocyte receptor modification was found to be a function of time and temperature. Titers of erythrocyte receptor-modifying activity were not diminished by successive exposures to fresh erythrocytes. Erythrocytes treated with erythrocyte receptor-modifying factor suspensions failed to significantly adsorb test virus hemagglutinin.

Inhibition of erythrocyte receptor modifying-activity of the adenovirus suspensions by rabbit antiserum was type-specific.
SELECTIVE ADRENAL NECROSIS AND APOPLEXY INDUCED BY 7,12-DIMETHYLBENZ(a)ANTHRACENE
Authors: Huggins Charles , Morii Sotokichi
Published: June 2004

Invariably in every normal rat a single dose of 7, 12-dimethylbenz(a)anthracene, by mouth or injected in a vein, was found to cause apoplexy and massive necrosis in the inner zones of the adrenal cortex; the zona glomerulosa, the adrenal medulla, and a small region of cortex adjacent to the great adrenal vessels were spared from damage. DMBA caused these selective lesions in females and in males of 2 strains of rats. Hemorrhage and necrosis were observed in no organ other than the adrenal gland. Whereas adrenal glands were heavily damaged by DMBA, pituitary and ovary escaped injury by the compound.

A single huge but sublethal feeding of o, p'-DDD caused degenerative changes of minor magnitude in the adrenals and only in a small percentage of rats; the property of inducing adrenal damage was not shared by other polynuclear aromatic hydrocarbons which were investigated, including strong carcinogens.

Presence of adrenal medulla is not a prerequisite to damage of the adrenal cortex by DMBA. The adrenal damage occurred in rats, given DMBA, from which the pituitary had been removed but the lesions were smaller in extent and less in incidence as post-hypophysectomy atrophy of the adrenal cortex progressed.

The entire DMBA molecule was necessary to induce adrenal damage; fragments of this molecule did not induce adrenal lesions. Two components which are of cardinal importance in this specific damaging effect are: (a) electronic factor; (b) steric factor.

The level of isocitric dehydrogenase in adrenal is modified considerably by presence or absence of estradiol-17ß.
THE APPLICATION OF FERRITIN-CONJUGATED ANTIBODY TO ELECTRON MICROSCOPIC STUDIES OF INFLUENZA VIRUS IN INFECTED CELLS
Authors: Morgan Councilman , Hsu Konrad C. , Rifkind Richard A. , Knox Alice W. , Rose Harry M.
Published: June 2004

Freezing of the chorioallantoic membrane after brief fixation in formalin preserves antigenicity of cellular components and allows penetration of ferritin-conjugated antibody. Dense aggregates of viral antigen, presumed to be of the soluble type, were found in the nuclei of cells infected with influenza virus. Intracytoplasmic antigen, on the other hand, was widely dispersed. The experimental observations are consistent with the hypothesis that soluble antigen diffuses into the cytoplasm through intact nuclear membranes. Nucleoli do not appear to be the sites of antigen synthesis.
OBSERVATIONS ON THE FINE STRUCTURE OF MATURE HERPES SIMPLEX VIRUS AND ON THE COMPOSITION OF ITS NUCLEOID
Authors: Epstein M. A.
Published: June 2004

The fine structure and composition of mature herpes virus have been investigated in thin sections by electron microscopy. The virus was grown in cultured HeLa cells and was collected with them. Tests for the biological activity of the infected cultures were included in the first half of the work. Preparations were fixed with both osmium and permanganate, and were embedded either in methacrylate or in aquon, a water-miscible fraction of a commercially available epoxy resin. In further experiments material fixed with permanganate was subjected to the action of specific nucleases or control medium before embedding.

All the preparations showed numerous uniform particles around and between the cells and this was paralleled by considerable biological activity where tests were made on samples of the culture fluids.

Mature herpes virus has been found to be round and to measure, when dehydration shrinkage is avoided, about 165 mµ in diameter. The particle contained an eccentric round-ended rod-shaped, electron-opaque nucleoid lying in an inner zone of low density. A dense outer zone or viroplasm surrounded this, no membrane being present between the two zones. After permanganate fixation the particle was found to have an outer limiting membrane showing a triple-layered structure morphologically indistinguishable from that of the plasma membrane of the HeLa cells.

The results of the digestion experiments show that herpes virus contains nucleic acid of deoxyribose type and that this is localized in the dense nucleoid.

Both the findings and the methods used are discussed.
CRITICAL RELATIONSHIPS BETWEEN CONSTITUENTS OF THE ANTIGEN-ADJUVANT EMULSION AFFECTING EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS IN A COMPLETELY SUSCEPTIBLE MOUSE GENOTYPE
Authors: Lee Johanna M. , Schneider Howard A.
Published: June 2004

The production of EAE in the fully susceptible BSVS mouse genotype has been found to be dependent on the ratio of proteolipid antigen and adjuvant mycobacterial concentration as used in the emulsion, of the Freund type. Disturbance of this ratio, by manipulation of either component, by diminution or increase, results in a decrease in the frequency by which EAE is produced.

Simultaneous reduction of antigen and mycobacteria, so that the ratio remains unchanged, retains the full EAE-producing power of the emulsion. The limit of this has not been ascertained.

Emulsifying agents have been found to restrict further the permissible limits of the antigen-mycobacterial ratio for full EAE production. Such effects of the emulsifier have been found to vary with the qualitative nature of the emulsifier. Aquaphor has been found to be less restrictive than falba.

These phenomena, systematically analyzed here for the mouse, may have an application for other antigen-adjuvant systems and for other hosts.
ADJUVANT-ANTIGEN RELATIONSHIPS IN THE PRODUCTION OF EXPERIMENTAL "ALLERGIC" ENCEPHALOMYELITIS IN THE GUINEA PIG
Authors: Shaw Cheng-Mei , Alvord Ellsworth C. , Fahlberg Willson J. , Kies Marian W.
Published: June 2004

Quantitative relationships between the amounts of encephalitogenic neural tissue and mycobacterial adjuvant are presented for the guinea pig, for comparison with similar results obtained in the mouse and presented in an accompanying paper by Lee and Schneider (27). Definite threshold amounts of both neural encephalitogenic and mycobacterial adjuvant can be defined. With the proper amounts of each component, 100 per cent of guinea pigs can be made to develop EAE, 97 per cent dying of it, and over 50 per cent becoming paralyzed by the 12th day after challenge. With moderate amounts of encephalitogen the severity and incidence of EAE can be very great, but this encephalitogenic potential can be masked if large amounts of mycobacteria are employed. The mechanism of this masking effect by excess adjuvant is not known, but speculation centers upon the possibility of competition of antigens of the mycobacteria at the expense of those of the encephalitogen.
AUTORADIOGRAPHIC STUDIES ON THE IMMUNE RESPONSE
Authors: Nossal G. J. V. , Mäkelä O.
Published: June 2004

The origin and growth kinetics of plasma cells have been investigated using autoradiographic labeling techniques. Rats immunized once with Salmonella flagella were given a single pulse of H3-thymidine 4 or 40 weeks later. 2 hours after the tracer injection, they received a secondary antigenic stimulus. When animals were sacrificed immediately only certain cells from the resting primarily immunized lymph nodes, notably large and medium lymphocytes, were labeled. Subsequent to secondary stimulation, animals were killed at intervals; nearly all the plasma cells formed within the next 5 to 6 days were labeled. They must thus have been the progeny of cells already capable of synthesizing DNA in resting nodes, most probably of large lymphocytes. Plasmacytopoiesis began with little or no lag following secondary immunization, and the number of labeled plasma cells rose exponentially between the 2nd and 4th day, with a doubling time of about 12 hours. Studies of mean grain counts of primitive cells also suggested that the generation time of plasmablasts was 12 hours or less. The hypothesis was proposed that immunological memory depended on the persistence, following primary stimulation, of a continuously dividing stem line of primitive lymphocytes, reactive at all times to further antigenic stimulation.
COMPARATIVE STUDIES IN ROUS SARCOMA WITH VIRUS, TUMOR CELLS, AND CHICK EMBRYO CELLS TRANSFORMED IN VITRO BY VIRUS
Authors: Dougherty Robert M. , Morgan Herbert R.
Published: June 2004

Chick embryo fibroblasts infected in vitro with Rous sarcoma virus have properties similar to tumor cells when injected into virus-immune chickens. When such virus-transformed fibroblasts are injected into normal chickens, they apparently participate in the production of tumors independent of their release of virus and are thus apparently malignant in vivo.
IMMUNITY AGAINST TULAREMIA: PASSIVE PROTECTION OF MICE BY TRANSFER OF IMMUNE TISSUES
Authors: Allen William Peter
Published: June 2004

Acquired resistance to fully virulent strains of P. tularensis was passively transferred to normal mice by viable spleen cells or peritoneal leucocytes from donors that recovered from infection with an attenuated strain of P. tularensis. This passively transferred resistance was reflected in survival or delayed death after challenge of the recipients. The degree of passively transferred resistance was dependent upon the immune status of the donors and the number of viable immune cells transferred. There were indications that this resistance persisted only as long as the transferred tissues were compatible with the tissues of the recipient. The results support the hypothesis that immunity of mice against fully virulent strains of P. tularensis is associated with an altered state of the tissues.
TRANSFER OF AN AUTOIMMUNE NEPHROSIS IN THE RAT BY MEANS OF LYMPH NODE CELLS
Authors: Hess Evelyn V. , Ashworth Charles T. , Ziff Morris
Published: June 2004

An autoimmune nephrosis produced in rats by repeated injections of kidney extract with Freund's adjuvant has been transferred by means of lymph node cells to recipient animals rendered tolerant by neonatal injection with spleen cells from prospective donors. Transfer of the disease was manifested in the recipients by the development of proteinuria, hypoalbuminemia, hypercholesterolemia,and histological changes. The latter consisted of glomerular epithelial swelling, increase in basement membrane material and the presence of protein droplets in the glomerular and tubular epithelium. Appropriate control experiments were negative. Attempts to transfer with serum were unsuccessful. The transfer described is believed to provide evidence for an immunological mechanism for kidney and adjuvant induced nephrosis in the rat.
THE CHEMOTACTIC EFFECT OF MIXTURES OF ANTIBODY AND ANTIGEN ON POLYMORPHONUCLEAR LEUCOCYTES
Authors: Boyden Stephen
Published: June 2004

An in vitro technique is described for assessing the chemotactic activity of soluble substances on motile cells.

Antibody-antigen mixtures when incubated (37°C) in medium containing fresh (i.e. non-inactivated) normal rabbit serum exert a strong chemotactic effect on rabbit polymorphonuclear leucocytes.

Results are described which indicate that, when antibody-antigen complexes are incubated (37°C) in fresh serum, a heat-stable (56°C) substance (or substances) is produced which acts directly as a chemotactic stimulus on the polymorphs. This heat-stable chemotactic substance is not produced when antibody-antigen complexes are incubated in serum which has been heated at 56°C for 30 minutes.
STUDIES ON TUBERCLE BACILLUS-HISTIOCYTE RELATIONSHIP
Authors: Fong Jacob , Chin Dennis , Elberg Sanford S.
Published: June 2004

Studies of passive transfer of cellular resistance, as manifested by refractoriness to necrotization with virulent tubercle bacilli, have shown that immune histiocytes or immune lymphocytes were effective transferring agents; immune polymorphonuclear leucocytes and immune serum as well as comparable cells from normal animals lacked this capacity.

Comparisons of immune histiocytes and immune lymphocytes showed that the former cells were more efficient; this was indicated by (a) the smaller numbers of immune histiocytes needed for passive transfer, (b) the longer duration of cellular resistance in recipients given histiocytes than in those given lymphocytes, (c) the greater capacity of histiocytes to effect serial passive transfer, and (d) the ability of histiocytic but not lymphocytic lysates to transfer cellular resistance.

Experiments to establish the mechanism of passive transfer of cellular resistance showed that there was no active induction of resistance in recipients through transfer of bacillary antigens contained in immune histiocytes; in fact, the results of serial passive transfers with immune histiocytes suggested an active replication of the "cell resistance factor."
CYTOMETRIC ASSAY OF TOXICITY OF BRUCELLA ANTIGENS FOR SENSITIZED AND NON-SENSITIZED CELLS FROM THE GUINEA PIG
Authors: Carpenter Charles M. , Fukuda Morimichi , Heiskell Charles L.
Published: June 2004

1. A significant cytotoxic effect of trypsinized, sensitized cell cultures was observed when re-exposed to Brucella antigens in vitro.

2. The cytometric assay of the toxic effects of Brucella antigens using trypsinized cells from sensitized animals provides an objective measure of induced cellular hypersensitivity.
THE EFFECT OF BACTERIAL ENDOTOXIN ON THE NEWBORN GUINEA PIG
Authors: Uhr Jonathan W.
Published: June 2004

Newborn guinea pigs (less than 24 hours old) are highly susceptible to the lethal effect of endotoxin and susceptibility decreases with age. The vulnerability to endotoxin has not been attributed to hypofunction of either the adrenal gland or the reticuloendothelial system of the newborn, since prior injection of corticosteroids provides only partial protection to endotoxin and newborn guinea pigs can efficiently remove colloidal substances from the circulation. In contrast to adults, newborns do not develop a delayed inflammatory reaction following intradermal injection of endotoxin, and after intravenous challenge frequently fail to display neutropenia and sustained fever.
THE ENHANCEMENT OF CHLOROFORM-INDUCED PLASMA PROTEOLYTIC ACTIVITY BY EPSILON AMINOCAPROIC ACID
Authors: Donaldson Virginia H. , Ratnoff Oscar D.
Published: June 2004

The proteolytic activity in chloroform-treated plasma euglobulins has been attributed to plasmin. Plasmin can digest both casein and fibrin. Epsilon aminocaproic acid, which inhibits the activation of plasminogen, the precursor of plasmin, by streptokinase, urokinase, and tissue activators enhanced the development of casein hydrolytic activity in a mixture of chloroform and plasma euglobulins. Fibrinolytic activity was also enhanced, but this was evident only if the epsilon aminocaproic acid was removed from the chloroform-treated euglobulins prior to assay. The reasons for the paradoxical enhancement of chloroform-induced casein hydrolysis by euglobulins containing epsilon aminocaproic acid are unclear. However, studies of optimal pH, heat stability, and the effect of ionic strength on the activation of the precursor of this proteolytic enzyme do not differentiate it from plasminogen.
AN ALUMINUM MARKER FOR THE DIFFERENTIATION AND SEPARATION OF VIRULENT AND ATTENUATED POLIOVIRUSES
Authors: Wallis Craig , Melnick Joseph L. , Ferry George D. , Wimberly Ira L.
Published: June 2004

A new character, the A marker, for polioviruses is described. In the presence of Al+++ (1 to 100 mM) attenuated, but not virulent, strains of type 1 and type 2 polioviruses are stabilized so that they resist thermal inactivation at 50°C. Differences between the attenuated and virulent type 3 strains studied were of the opposite character and less marked.

By the use of optimal concentrations of Al+++ (12 mM for type 1 and 1 mM for type 2) attenuated strains can be separated from mixtures containing virulent type 1 or 2 strains.

The A marker was found to be correlated with the d but not the rct/40 or T marker. The A marker has been used for following genetic changes of vaccine strains after their multiplication in man.
IMMUNOLOGICAL SPECIFICITY OF DELAYED AND IMMEDIATE HYPERSENSITIVITY REACTIONS
Authors: Benacerraf B. , Levine B. B.
Published: June 2004

The effects of the following parameters on the immunologic specificity of delayed and immediate hypersensitivity reactions were investigated in the guinea pig using the picryl and p-toluenesulfonyl systems: (a) the contribution of the carrier protein, (b) the effect of the number of hapten groups per molecule of the immunizing and challenging antigens, and (c) the effect of interposing a 6 carbon chain (ϵ-aminocaproic acid) between the hapten and its usual attachment to the lysine ϵ-NH2 groups of the carrier protein.

It was found that induction of delayed hypersensitivity was accomplished equally well with both lightly and heavily coupled conjugates. Sensitized animals which gave strong delayed reactions to the immunizing conjugate cross-reacted poorly or not at all to (a) conjugates of the same hapten with a different carrier protein, or (b) conjugates differing from the immunizing conjugate by having an ϵ-aminocaproyl chain interposed between hapten and its attachment onto the carrier protein. Animals sensitized with either lightly or heavily substituted conjugates exhibited strong delayed reactions to both conjugates, but more intense reactions to the immunizing conjugate were always observed.

In contrast to the marker carrier specificity exhibited by the delayed hypersensitivity reactions, immediate hypersensitivity reactions, (specific precipitation, Arthus, and PCA reactions) could be elicited equally well with hapten conjugates of all carrier proteins, as well as with conjugates containing ϵ-aminocaproyl chains interposed between hapten and the carrier protein, provided the number of hapten groups per molecule conjugate was sufficiently high. Both in inducing antibody response and in provoking immediate hypersensitivity reactions, heavily substituted conjugates were considerably more effective than were lightly substituted conjugates.

Alternative explanations for these observed differences in specificity between immediate and delayed hypersensitivity reactions are discussed.
DELAYED HYPERSENSITIVITY TO HAPTEN-PROTEIN CONJUGATES
Authors: Cell P. G. H. , Silverstein Arthur M.
Published: June 2004

Further data have been presented showing that the specificity of the delayed hypersensitivity reaction in the guinea pig to hapten-protein conjugates involves to a considerable degree a contribution by the protein carrier. The carrier contribution is such that sensitization to guinea pig albumin-m-azobenzenesulfonate, for example, does not result in cross-reaction with conjugates of the same hapten with unrelated proteins such as ovalbumin or human gamma globulin, nor were cross-reactions observed between conjugates prepared with the same hapten, coupled to the same protein, but by two different chemical routes, such that the point of attachment of the hapten to the protein differed. It thus appears that in this system both hapten and carrier protein are necessary, but that neither alone is in general sufficient to stimulate the delayed sensitive cell.

Desensitization experiments with cross-reacting hapten-protein conjugates have suggested the presence of a multiplicity of antigenic determinants participating in the elicitation of the delayed lesion, and of a concomitant development of a heterogeneity of specificities in the population of delayed sensitive cells in the sensitized animal.

The data are discussed in terms of the apparent requirement of the delayed sensitivity mechanism for a larger functional antigenic determinant than that required for interaction with circulating antibodies. Some possible explanations for this difference, and some of its consequences, are discussed.
IMMUNOHISTOLOGIC STUDIES ON ANTIGEN-ANTIBODY REACTIONS IN THE AVASCULAR CORNEA
Authors: Germuth Frederick G. , Maumenee A. Edward , Senterfit Laurence B. , Pollack Abou D.
Published: June 2004

The injection of antigen into the center of the avascular cornea of homologously sensitized animals induced a ring of opacification between the center of the cornea and the limbus. This ring of opacification was composed of a line of deeply eosinophilic amorphous material in a matrix of swollen collagen fibers, palisaded by polymorphonuclear leucocytes. By use of fluor-tagged antigen, it was shown that the line of damage in the cornea coincided with the precipitation of antigen presumably by antibody entering the cornea from the limbal vessels. With the passage of time, the antigen-antibody precipitates were removed, at least in part by phagocytosis, and the ring of opacification was replaced by ingrowing blood vessels surrounded by plasma cells.

Treatment of sensitized animals with nitrogen mustard showed that antigen-antibody interaction could injure the corneal stroma in the relative absence of polymorphonuclear leucocytes.
THE HISTOLOGICAL DISTRIBUTION OF THE BLOOD GROUP SUBSTANCES IN MAN AS DISCLOSED BY IMMUNOFLUORESCENCE
Authors: Szulman Aron E.
Published: June 2004

The H antigen was mapped out by immunofluorescence in human tissues (including those of fetuses from 15 cm crown-heel length) from individuals of the various groups within the ABO system, both secretors and non-secretors. The distribution of the antigen can be summarized under the following headings:

Cell walls of endothelium: present throughout the cardiovascular system;

Cell walls of stratified epithelia: in skin, non-cornifying squamous stratified membranes, transitional epithelia;

Mucus: occurring wherever the latter is produced in secretor individuals and confined to a few special topographical areas in non-secretors;

Secretions and excretions: the pancreatic and sudoriferous (independent of secretor status), and mammary and uterine (governed by the secretor makeup) all contain it.

The distribution of the H antigen is most fully represented in tissues of group O. It follows an over-all universal pattern, characteristically modified in non-secretors, equally valid for antigens A and B described in a preceding study. Within this pattern, in tissues of the non-O groups, the complement of the H substance in its various forms wanes in a manner consistent with the hypothesis that it serves as a substrate for the A1, A2, B genes, exerting their action with different degrees of efficiency.

The secretor:non-secretor phenomena can be most simply interpreted by viewing the non-secretor, recessive gene (in the homozygous, ss condition) as inhibiting the production of some of the water-soluble forms of the blood group substances. Since the gene was never found responsible for dissociation of the H and A, B antigens its inhibitory action is thought to be wrought at the point of formation of the basic H substance or its precursor.
THE NATURE OF BENCE-JONES PROTEINS
Authors: Delman G. M. , Gally J. A.
Published: June 2004

The chemical relations among Bence-Jones proteins, myeloma proteins, and normal γ-globulins have been investigated by a variety of means. Starch gel electrophoresis in 8 M urea of reduced alkylated Bence-Jones proteins yielded patterns of bands corresponding to those of the light (L) polypeptide chains of the dissociated myeloma protein from the same patient.

One instance in which this correspondence was found was chosen for extensive study. Chromatography on carboxymethylcellulose in 6 M urea was employed to isolate the light (L) polypeptide chains and heavy (H) polypeptide chains of the completely reduced and alkylated myeloma protein. Isolation of similarly treated Bence-Jones protein from the same patient corroborated the correspondence to the L chains of the myeloma protein. Amino acid analyses indicated that the compositions of the Bence-Jones protein and the L chains of the myeloma protein were identical. Moreover, the thermosolubility properties and spectrofluorometric behavior of the isolated L chains and Bence-Jones protein were similar.

Ultracentrifugal analyses of the L chains of normal human 7S γ-globulin showed that their molecular weight in 6 M urea was 20,000. In aqueous solution their molecular weight was 41,000, suggesting that they exist as dimers under these conditions. The L chains of normal human γ-globulin were found to have reversible thermosolubility properties similar to those of Bence-Jones proteins. The H chains of normal human γ-globulin did not share these properties.

Using spectrofluorometric methods, characteristic molecular transitions were found upon heating Bence-Jones proteins and L chains. These transitions were indicated by an increase in the intensity of fluorescence at well defined temperatures as well as by reversible shifts in the wavelength of maximal emission.

The findings suggest that Bence-Jones proteins are composed of L chains of the type found in normal and pathological γ-globu]ins.
THE EFFECT OF HYDROCORTISONE ON THE RESPONSE OF FETAL RAT SKIN IN CULTURE TO ULTRAVIOLET IRRADIATION
Authors: Weissmann Gerald , Fell Honor B.
Published: June 2004

1. The effect of hydrocortisone on the development of fetal rat skin in organ culture, and on its repair after exposure to a mixed beam from a mercury lamp, are described.

2. The addition of hydrocortisone (7.5 µg/ml) to the culture medium (HC medium) caused accelerated differentiation and keratinisation of the epidermis followed by atrophic changes as in vivo.

3. Explants were grown for 2 days in either normal or HC medium and then irradiated with light from an Hanovia lamp.

4. Irradiation of the control explants produced severe necrosis in both epidermis and dermis and much disorganisation of the dermal intercellular material; 2 days after exposure the s. corneum with adherent cellular debris had become either completely detached from the denuded dermis, or raised to form a fluid-filled blister. Epidermal regeneration had begun by the 4th day after irradiation and was usually complete by the 6th day.

5. Hydrocortisone modified the response to irradiation as follows: (1) reduced and retarded cellular breakdown, (2) prevented vesication, (3) preserved the organisation of the dermal intercellular material, (4) hastened epithelialisation, (5) accelerated the differentiation of the new epidermis. Effects (2), (3), and (4) were probably secondary to (1).

6. Experiments with various light filters showed that the effective wavelengths for producing lesions in the skin explants were those below 3000 A.

7. It is suggested that the beneficial effect of hydrocortisone on the repair of irradiated skin explants might be due, at least in part, to a reduced proteolytic activity in the damaged tissue through a stabilising action of the hormone on the lysosomes.

8. The relationship of these findings to clinical observations is discussed.
INTRACELLULAR LOCALIZATION AND QUANTITATION OF TRITIATED ANTIGENS IN RETICULOENDOTHELIAL TISSUES OF MICE DURING SECONDARY AND HYPERIMMUNE RESPONSES
Authors: Roberts Audrey N. , Haurowitz Felix
Published: June 2004

Autoradiography and quantitative radiochemical techniques have been used to determine intracellular localization of tritium and the quantity of tissue-bound tritium, respectively, following injections of H3-aniline azo PGG or H3-arsanilazo PGG to yield hyperimmune or secondary response stimulation in mice. Autoradiography revealed intracytoplasmic localization of grains in macrophages of spleen and lung sections, and in Kupffer cells of liver sections following intravenous and subcutaneous injections of H3-aniline azo PGG. Quantitation of tissue section surface radioactivities in the windowless flow counter and scintillation counter, and of dissolved tissue section activities in the scintillation counter, showed that greatest radioactivity was present in lung tissue, with less in spleen, liver, and mesenteric lymph nodes from these hyperimmunized mice.

Autoradiographic studies on tissue sections from mice in secondary response stimulation after subcutaneous foot-pad injections of H3-arsanilazo PGG, showed intracellular and extracellular grains over regional popliteal node sections, with intracytoplasmic grain localization over macrophages and pyroninophilic plasmacytes. Scattered macrophages in spleen and lung sections also contained intracytoplasmic radioactivity. Clusters of antibody-synthesizing cells in the regional lymph nodes were demonstrated with fluorescence microscopy, and these cells were compared to similar cells possessing radioactivity as observed in the section autoradiographs. An occasional Russell body plasma cell containing specific antibody was observed in splenic impressions. Windowless flow counting showed that greatest radioactivity was in regional node sections, with less in spleen and lung, and none in contralateral lymph nodes.

A quantitative comparison between windowless flow counting and autoradiography revealed that 20 counts were required to yield one silver grain.
STUDIES ON LYSOSOMES
Authors: Weissmann Gerald , Thomas Lewis
Published: June 2004

Granular fractions sedimenting between 800 g and 15,000 g have been prepared in 0.25 M sucrose from the livers of normal young rabbits and from the livers of rabbits injected with A. aerogenes endotoxin. As early as 5 minutes after endotoxin, there was an augmented release of two enzymes, beta glucuronidase and cathepsin, into the supernatant of fractions which had been incubated or incubated and irradiated with a mercury vapor lamp at 37°C. These effects were maximum at 30 minutes after endotoxin and were associated with a depletion of the total activities of the two enzymes within the granules. Concurrently there was a rise in the activity of the two enzymes in fractions of the homogenate which were unsedimentable at 15,000 g. Fractions prepared from animals made tolerant to endotoxin no longer responded to incubation and irradiation by an augmented release of these two hydrolases, nor did fractions prepared from animals pretreated with glucocorticoids. Pretreatment with DOCA did not prevent release of enzymes after endotoxin.

The results have been interpreted to indicate that one consequence of the injection of endotoxin is an effect upon the stability of lysosomes, with subsequent release of acid hydrolases into the cell sap or surrounding tissue. Glucocorticoids may exert their pharmacologic effects through the protection of these subcellular particles against a variety of injurious agents.
ANTIGENICITY OF POLYPEPTIDES (POLY ALPHA AMINO ACIDS)
Authors: Maurer Paul H. , Gerulat Bernard F. , Pinchuck Paul
Published: June 2004

It has been shown that the polymers GLA5, GLT, GLA40, and GLAT are antigenic in human beings. Both immediate skin reactions and delayed (cellular) reactions were observed.

The antibody produced reacted well in the precipitin reaction, agglutinated antigen coated tanned sheep cells, was less effective in inducing PCA reactions, and failed to sensitize guinea pigs for passive systemic anaphylaxis.

The immediate skin reactivity appeared not to be related to the P-K antibody. The delayed reaction was histologically distinct from the immediate reaction and consisted of perivascular lymphocytic infiltration in contrast to the predominantly polymorphonuclear cells in the latter reaction. The antigenic response against the polymers resembled that observed with proteins. From cross-reactions it appears that the response is heterogeneous in the sense that not all individuals react against the same groupings in the complex polymers.
STRUCTURE AND DEVELOPMENT OF VIRUSES AS OBSERVED IN THE ELECTRON MICROSCOPE
Authors: Morgan Councilman , Hsu Konrad C. , Rose Harry M.
Published: June 2004

Chicken embryos were infected by the chorioallantoic route with influenza virus, PR8 strain, in the form of undiluted chorioallantoic fluid. Electron microscopic examination 24 hours after infection revealed that membrane-bound fragments of cytoplasm appeared to be in process of release from entodermal cells of the chorioallantois. The number of such fragments was greatly increased in proportion to the number of typical viral particles after the third serial passage, which was accompanied by a reduction of the infectivity-hemagglutinin ratio (von Magnus effect). The lack of recognizable internal components, together with the presence of surface structure which closely resembled that of the virus and frequently contained viral antigen, suggested that many of these fragments were incomplete viral particles. It is proposed that concentrated inocula damage the cells and interfere with differentiation of the virus, but do not inhibit formation and detachment of cytoplasmic processes. Under these circumstances the accumulation of viral antigen at the surface of the cell will result in the predominant formation of incomplete virus.
IMMUNOHISTOCHEMICAL DEMONSTRATION OF THE RETICULOENDOTHELIAL CLEARANCE OF CIRCULATING FIBRIN AGGREGATES
Authors: Lee Leung , McCluskey Robert T.
Published: June 2004

In rabbits given an intravenous infusion of thrombin or an injection of endotoxin, immunohistochemical examination of the tissues, using conjugated antiserum against rabbit fibrin, showed bright intracytoplasmic staining in many of the phagocytic cells of the liver and spleen. In normal rabbits as well as in animals injected with large doses of heparin prior to thrombin or endotoxin administration, no such intracellular staining was observed. The findings of this study substantiate the hypothesis that fibrin aggregates formed in the circulating blood during low grade intravascular coagulation are largely removed by the reticuloendothelial system.
THE EFFECT OF 6-MERCAPTOPURINE ON DELAYED HYPERSENSITIVITY IN GUINEA PIGS
Authors: Hoyer John R. , Hoyer Leon W. , Good Robert A. , Condie Richard M.
Published: June 2004

The development of tuberculin hypersensitivity in guinea pigs after BCG stimulation was suppressed by intramuscular administration of 50 mg/kg/day of 6-mercaptopurine started at the time of stimulation. Fasting of guinea pigs after BCG stimulation had no effect on the development of tuberculin hypersensitivity.
SIGNIFICANCE OF CRYOPROFIBRIN IN FIBRINOGEN-FIBRIN CONVERSION
Authors: Shainoff John R. , Page Irvine H.
Published: June 2004

Fibrinogen altered by thrombin-catalyzed liberation of fibrinopeptide A was found to combine with native fibrinogen to form a cold-precipitable complex we have called "cryoprofibrin." The altered fibrinogen lacking fibrino-peptide A polymerized into fibrin, but not until conditions for equilibrium between its incorporation into both cryoprofibrin and fibrin were satisfied. At equilibrium, the concentration of cryoprofibrin was maintained at a threshold proportional to the concentration of fibrinogen. When the concentration of cryoprofibrin was below threshold, fibrin could be depolymerized and solubilized by fibrinogen with resultant formation of cryoprofibrin. Since threshold concentrations of cryoprofibrin appear necessary for precipitation of fibrin, the concentration of cryoprofibrin in plasma provides a basis for determining intravascular deposition of fibrin. Intravascular deposition of fibrin does not appear to occur normally in rabbits, because the concentration of cryoprofibrin in plasma from normal rabbits is far below the threshold for precipitation of fibrin. The applicability of cryoprofibrin as an indicator of fibrin deposition is demonstrated by the occurrence of levels of cryoprofibrin approaching the threshold for precipitation of fibrin in plasma from endotoxin-treated rabbits.

The current concept that the fibrinogen molecule can dissociate into subunits can be used to explain the conversion of fibrinogen to cryoprofibrin. As one possibility, the two residues of fibrinopeptide A contained in fibrinogen may be located on two separate subunits of the molecule; cryoprofibrin is produced when one of these subunits is replaced by a subunit altered by loss of fibrinopeptide A. Recombination of native subunits with subunits altered by loss of A would counter dissociation of cryoprofibrin and inhibit polymerization of subunits lacking fibrinopeptide A. As an alternate mechanism, two residues of A may be liberated concurrently from a single subunit. Cryoprofibrin would then correspond to a fibrinogen molecule, containing a subunit with two residues of A, in combination with an altered molecule containing a subunit lacking two residues of A.

Liberation of fibrinopeptide B did not contribute measurably to production of fibrin resulting from limited action of thrombin on rabbit fibrinogen. Both fibrin containing B but not A, and fibrin containing neither B nor A, as is produced by extensive action of thrombin, could be solubilized by fibrinogen.

Thrombin, or another enzyme utilizing tosyl-L-arginine methyl ester as substrate, appeared reversibly to inhibit polymerization of fibrin containing fibrinopeptide B. This enzyme and fibrinogen were the only proteins appearing to inhibit polymerization in plasma from normal rabbits.
ANTIBODY STUDIES IN RABBIT ENCEPHALOMYELITIS INDUCED BY A WATER-SOLUBLE PROTEIN FRACTION OF RABBIT CORD
Authors: Kibler Robert F. , Barnes Arthur E.
Published: June 2004

A water-soluble protein fraction of nervous tissue was prepared by extraction of rabbit spinal cord with sodium citrate at pH 4.3. Characterization by nitrogen content and paper electrophoresis showed it to be a mixture of basic proteins. This extract demonstrated encephalitogenic activity when injected into rabbits.

The most suitable technique for the measurement of serum antibody to the rabbit cord antigen proved to be the precipitation of antigen-antibody complexes by 40 per cent saturated ammonium sulfate. Antibody could not be demonstrated by the techniques of complement fixation, quantitative precipitation, and Ouchterlony plates.

The early appearance of circulating antibody occurred almost exclusively in rabbits that subsequently developed EAE. Specificity of the antibodies for nervous tissue was demonstrated by appropriate blocking and adsorption experiments.
MEASUREMENT OF RESISTANCE TO EXPERIMENTAL TUBERCULOSIS IN MICE
Authors: Costello Richard , Dubos Rene , Schaedler Russell W.
Published: June 2004

NCS mice were found to be susceptible to a hyperacute form of disease induced by injection of relatively large doses of Mycobacterium tuberculosis. Susceptibility to this type of disease was conditioned by the sex, age, and dietary history of the animals. Unlike the more usual response of mice to mycobacterial disease, these conditions affected primarily the relative occurrence of early-acute deaths rather than average survival time.

Resistance to this form of disease was increased by previous exposure of the animals to Gram-negative bacteria, to their endotoxin, or to various preparations of killed mycobacterial cells.
THE ROLE OF HUMORAL ANTIBODIES IN REJECTION OF SKIN HOMOGRAFTS IN RABBITS
Authors: Kretschmer Roberto R. , Pérez-Tamayo Ruy
Published: June 2004

Passive transfer of transplantation immunity by means of sensitized lymphoid cells enclosed within diffusion chambers has been accomplished in non-radiated and radiated rabbits. This result, together with other data available in the literature, suggests that humoral antibodies play an important role in rejection of skin homografts in the rabbit.