bluuebunny/crossref_metadata_2025 huggingface dataset:
INACTIVATION OF BACTERIAL EXOTOXINS AND ENDOTOXIN BY IRON
Authors: Janoff Aaron , Zweifach Benjamin W.
Published: June 2004

After incubation with various agents in vitro, the lethal effects of aqueous solutions of Cl. perfringens alpha toxin, C. diphtheriae toxin, and E. coli endotoxin were tested in mice, guinea pigs, and rabbits, respectively.

Iron, copper, cysteine, ascorbic acid, and versene counteracted the lethal effects of alpha toxin in mice, while magnesium, manganese, zinc, and citrate did not.

Iron also counteracted the lethal effects of diphtheria toxin in guinea pigs. After incubation of endotoxin with iron, its lethal effects and tissue-necrotizing actions in rabbits were counteracted. However, the pyrogenic properties of the toxin were not affected.

The solubilities of perfringens alpha toxin and diphtheria toxin were markedly reduced after incubation with detoxifying metals, and resolubilization of these toxins with chelators resulted in partial restoration of toxicity. Addition of versene to detoxified endotoxin also resulted in partial recovery of lethal effectiveness.

The inactivation of bacterial toxins by iron under in vitro conditions is not specific to this metal, is a reversible process, and may be due to desolubilization, reduction, or to competition by the metal for sites on the toxin normally bound by other cations in vivo.

Although no evidence is presented in this paper to support the view that there is a relationship between the inactivation of endotoxin and the storage iron in the reticuloendothelial system of shocked animals, the observation of an in vitro inactivation of endotoxin by inorganic iron warrants consideration of such a mechanism.
DEPRESSION BY ANTIBODY OF THE IMMUNE RESPONSE TO HOMOGRAFTS AND ITS ROLE IN IMMUNOLOGICAL ENHANCEMENT
Authors: Snell George D. , Winn Henry J. , Stimpfling Jack H. , Parker Stephen J.
Published: June 2004

This paper reports tests of two hypotheses that have been proposed to account for the enhanced growth of tumor homografts in the presence of antiserum reactive with the graft (immunological enhancement). According to the first hypothesis, enhancement is due to some "physiological" alteration in the tumor, induced by its contact with antiserum, which insures its survival despite the hostile response of the host. According to the second hypothesis, antiserum alters the response of the host. By blocking the development of the cellular type of immunity, which is the main agent in graft destruction, it permits the graft to survive.

To test hypothesis 1, strain A tumor SaI was passed from A's, and from enhanced B10.D2's, into untreated B10.D2's. The per cent of deaths was essentially the same in both groups (48 and 44 per cent, respectively); there was no evidence that passage through enhanced B10.D2's altered the capacity of the tumor to grow in the foreign strain. Several other groups of mice included in the experiment all confirmed this conclusion. The experiment failed to confirm hypothesis 1.

In the tests of hypothesis 2, the effect of isoantiserum on immune responses of both the humoral and cellular type was measured. When antiserum was given together with foreign strain lymphoid cells (antigen), almost no additional antibody was manufactured; in contrast with this, controls receiving foreign cells only produced red cell agglutinating antibody in high titer.

The effect of antiserum on the development of immunity of the cellular type was tested by the method of Winn. In this assay, presumptively immune node cells, in various dilutions, are mixed with tumor cells and injected into appropriate mice. Immunity is indicated by inhibited tumor growth. Antiserum given at the same time as a tumor homograft greatly depressed the immunity of the cells expressed from the draining nodes. At 6 days after the graft, the level of immunity of cells from treated mice was 1/24th to 1/32nd that of cells from controls receiving tumor alone. The same sort of depressing effect was noted when the immunizing tissue was foreign thymus or embryo. Antiserum given 1 or more days after the immunizing tissue also resulted in a lower level of cellular immunity (but the assay used in this case was a less critical one).

These results provide an adequate explanation of the phenomenon of immunological enhancement, at least as it occurs in the particular test system used in these experiments. Since it is cellular immunity rather than humoral antibody that inhibits the growth of most grafts (transplantable leukemias are an exception), the depression of this immunity by antibody is favorable to the growth of a homograft.
DIPLOID HETEROZYGOUS HYBRIDS FROM MATINGS BETWEEN ESCHERICHIA COLI AND SALMONELLA TYPHOSA
Authors: Baron L. S. , Spilman Walter M. , Carey Warren F.
Published: June 2004

An Hfr strain of E. coli K-12 has been shown to mate at low frequency with a number of strains of S. typhosa. The hybrids, selected as lactose positives, retained all other antigenic and biochemical manifestations of the S. typhosa parent. A Lac+ hybrid, S. typhosa strain 643L+ was remated with the E. coli Hfr and plated on minimal media containing L-arabinose, D-xylose, L-rhamnose, or L-fucose as the sole carbon sources. Hybrids of the remated strain appeared at high frequency on the plates containing L-arabinose, and could be detected at lower frequencies on plates containing D-xylose, L-rhamnose, and L-fucose. Those selected for D-xylose or L-rhamnose utilization possessed the attributes of segregating diploid heterozygotes being highly unstable and continually segregating a cultural form typical of the S. typhosa parent. The unstable type exhibited most of the biochemical characteristics of the E. coli parent including the ability to produce indol, and also reacted with antiserum to both the E. coli and S. typhosa parent strains, owing to the acquisition of a thermolabile antigen from the E. coli parent. The parent and hybrid strains were examined in detail for changes in patterns of phage susceptibility and virulence. Acquisition of susceptibility to a number of the T phages, a characteristic of the E. coli parent, was observed in one of the hybrid types. A decrease in virulence of the diploid hybrid form of S. typhosa in the mouse virulence test was found.
GENETICS OF HUMAN CELL LINES
Authors: Djordjevic B. , Szybalski Waclaw
Published: June 2004

The human cell line D98S can be cultivated indefinitely in the presence of up to 3 x 10–5 M 5-bromodeoxyuridine (BUDR), without loss of cell viability. During this time, BUDR is incorporated into both strands of the DNA molecules, replacing up to 45 per cent of the thymidine and thereby rendering the cells highly sensitive to UV light and to x-rays. Cells grown for a limited period of time in the presence of 5-iododeoxyuridine (IUDR) become UV-sensitized, while prolonged cultivation with IUDR results in the loss of cell viability. The properties of the BUDR label permitted the demonstration that: (a) human DNA replicates in a "semiconservative" manner; (b) the degree of radiosensitization of BUDR-treated cells depends on whether the DNA has been substituted in one strand only ("unifilarly") or in both strands ("bifilarly"); (c) functional human DNA is produced during partial inhibition of protein synthesis. The potential applicability of this new rational principle of radiosensitization to the radiotherapy of neoplastic diseases is discussed.
THE DISTRIBUTION OF S35-LABELED BOVINE SERUM ALBUMIN IN NEWBORN AND IMMUNOLOGICALLY TOLERANT ADULT RABBITS
Authors: Garvey Justine S. , Eitzman Donald V. , Smith Richard T.
Published: June 2004

The fate of injected S35-labeled sulfanilic acid-azoalbumin in the serum, various organs, and liver fractions was compared in the newborn and adult rabbit and in specifically unresponsive and normal adult rabbits. Exponential decay of the injected antigen without the usual immune phase of elimination was observed in newborn and unresponsive adult animals. Comparison of organ distribution of radioactivity in adults and animals injected at birth and 21 days of age showed persistence in the liver at least as long as 3 weeks in all groups (which was the time chosen for observation). The slight differences in spleen and thymus concentrations with age are of undetermined significance. Comparison of the organ distribution of antigen in unresponsive adult rabbits and in normal ones showed slight differences which were similar to those predicted from previous immunization of adults which gave low grade antibody response. There was a slight selective accumulation of antigen in the nuclear fraction of liver homogenates of unresponsive animals, but no other differences were observed.
THE NATURE OF STORAGE IRON IN IDIOPATHIC HEMOCHROMATOSIS AND IN HEMOSIDEROSIS
Authors: Richter Goetz W.
Published: June 2004

Using three different methods of cells fractionation, hemosiderin granules were isolated from tissues (liver and/or spleen) of three patients. The samples were obtained from a case of idiopathic hemochromatosis, a case of thalassemia major with secondary (transfusional?) hemosiderosis, and a case of transfusional hemosiderosis associated with an unclassified anemia.

Iron, nitrogen, and protein content of the hemosiderin granules varied over a wide range. Electron microscopy of sectioned granules revealed aggregates of dense particles of different shapes, with diameters ranging from 10 A to about 75 A. In some of the granules dense particles corresponding to the iron hydroxide micelles of ferritin molecules were abundant. But many of the granules contained very few of these molecules.

The presence of ferritin and apoferritin in the samples of hemosiderin granules was demonstrated by means of precipitin tests in agar-gel, using rabbit antiferritin sera with known antibody nitrogen concentrations. At least three antigenic components were detected in highly purified crystalline ferritin prepared from tissues of the three patients; the hemosiderin granules contained the same antigens, but probably in much smaller quantities. Both ferritin and apoferritin molecules were extracted from hemosiderin granules, and were demonstrated in the electron microscope after suitable preparation.

The solubility curve of human ferritin in solutions of (NH4)2SO4 was investigated. The results indicate that substantial quantities of ferritin or apoferritin can be lost in saline, aqueous media during isolation of hemosiderin granules from cells.

It was shown by means of electron microdiffraction on selected hemosiderin granules that the dense particles represent forms of partly hydrated α-Fe2O3. The conditions necessary for electron microdiffraction in an electron microscope precluded an exact determination of the state of hydration of the α-Fe2O3 or of its structural relation to (FeOOH) micelles of pure ferritin in its undenatured state.

The findings were considered in the light of evidence on the structure and disposition of hemosiderin in situ in cells, and on the structure of ferritin. Differences between endogenous hemosiderin and hemosiderin derived from injections of colloidal iron compounds were pointed out. The evidence indicates that in hemochromatosis and in secondary hemosiderosis much of the inorganic storage iron in liver and spleen is derived from degraded ferritin.

The findings suggest that an abnormal cellular metabolic pathway of ferritin is implicated in the pathogenesis of hemochromatosis and transfusional hemosiderosis.
THE MAMMALIAN CELL-VIRUS RELATIONSHIP
Authors: McLaren Leroy C. , Holland John J. , Syverton Jerome T.
Published: June 2004

All primary cell cultures tested from numerous human and rhesus monkey tissues were susceptible to Coxsackie A9 virus infection and were found to contain a specific A9 virus receptor. Ability of these cells to adsorb actively virus correlated with the presence of this receptor. Attachment of A9 virus to cell receptor was specifically enhanced by calcium and magnesium ions. Human cells established in continuous culture from pretested susceptible cultures were found to have lost A9 virus susceptibility and A9 receptors but not receptors for adsorption of type 1 poliovirus or B1, B3, B5 Coxsackie viruses. These cultures were able also to produce infectious A9 virus after exposure to viral RNA. Coxsackie A9 virus as well as type 1 poliovirus, inactivated following reaction with debris prepared from susceptible cells, could be recovered fully in infectious form by treatment of the inactive systems at low pH. Recovery of infectious A9 virus was also possible with a chelating compound. Coxsackie A9 virus could also be dissociated by means of low pH after adsorption onto susceptible cells for 2 minutes at 1°C., but not after 1 hour incubation at 37°C., indicating that viral host cell activity may be required in order for irreversible enterovirus attachment to take place.
FURTHER STUDIES ON PASSIVE TRANSFER OF TOLERANCE TO PYROGENICITY OF BACTERIAL ENDOTOXIN
Authors: Freedman Henry H.
Published: June 2004

The effect of various schedules for inducing tolerance to bacterial endotoxin in donor rabbits upon suitability for demonstration of passive transfer of tolerance to pyrogenicity in normal recipients has been investigated. Long-term treatment of donors, through 5 weeks, is no more effective than a brief series of injections, adding further evidence that tolerance is not attributable to specific antibody to the endotoxin. Qualitative differentiation of the febrile pattern of passively tolerant recipients from that seen in control animals depends upon the magnitude of the test dose of pyrogen.

Passively tolerant rabbits respond to endotoxin with an acute leucopenia equivalent to that seen in controls suffering a full biphasic fever. Animals given daily injections of endotoxin continue to show the acute leucopenia, despite the early modification of the course of fever characteristic of endotoxin tolerance. The assumption that the leucopenia reflects damage to the leucocytes, with release of endogenous pyrogen, is not consistent with these findings. Rabbits rendered leucopenic by nitrogen mustard and then given endotoxin exhibit a rapidly developing fever of greater than normal intensity, the exaggeration of the febrile response being proportional to the severity of the induced leucopenia.

The implications of these findings for the pathogenesis of endotoxin-induced fever are discussed. The evidence supports the hypothesis that endotoxin produces fever by direct action rather than by release of endogenous leucocytic pyrogen. It is postulated that the lesser fever, in animals having normal numbers of circulating leucocytes, reflects a limitation of available endotoxin by the known rapid sequestration in the white blood cells at the time of the acute leucopenia. It is further suggested that the biphasic febrile response of the normal rabbit results from reinoculation of the blood stream by the temporarily sequestered endotoxin, the RES of the tolerant animal clearing the released endotoxin at a rate sufficient to prevent triggering the second phase of fever.
FACTORS AFFECTING THE CHAIN LENGTH OF GROUP A STREPTOCOCCI
Authors: Ekstedt Richard D. , Stollerman Gene H.
Published: June 2004

Minute amounts of M protein were detected in culture supernates of virulent Group A streptococci by type-specific inhibition of the long chain and the bactericidal tests for anti-M antibody. The amount of M protein that was detected by the inhibition of these biological systems was less than could be demonstrated by precipitation tests.

All strains of streptococci rich in M protein which were studied formed long chains when grown in sufficient concentrations of anti-M antibody. Very low concentrations of anti-M antibody escaped detection by the long chain test when strains of excessive M protein content were employed. Under such conditions the bactericidal test detected anti-M antibody more sensitively than the long chain test owing to the smaller inoculum employed in the former method.

The scission of streptococcal chains may be inhibited by union of antibodies with surface antigens other than M protein. Long chains were formed when M-negative, R-positive strains were grown in sera containing anti-R antibody.
METABOLISM OF ACID MUCOPOLYSACCHARIDES IN THE SHWARTZMAN PHENOMENON
Authors: Berenson Gerald S. , Dalferes Edward R.
Published: June 2004

The effect of the Shwartzman phenomenon on the metabolism of the acid mucopolysaccharides (MPS), hyaluronic acid and chondroitin sulfate, from rabbits with localized lesions was studied. Quantitative changes of MPS were observed by two techniques—the in vivo incorporation of glucose C14 into the MPS and the in vitro dilution of C14-labeled MPS. The results indicate an increased concentration of both MPS fractions in the skin lesions and suggest an increased biosynthesis of the compounds at the site of the lesions. Deposition of chondroitin sulfate from blood may have contributed in part to the concentration of this substance in the lesions. The MPS contribute to the inflammatory state of the Shwartzman phenomenon but the specific roles of these compounds need to be defined by further study.
INCREASED FIXATION OF SULFUR35 BY CARTILAGE IN VITRO FOLLOWING DEPLETION OF THE MATRIX BY INTRAVENOUS PAPAIN
Authors: McElligott T. F. , Potter J. L.
Published: June 2004

The uptake in vitro of sulfur-35 by costal cartilage obtained from nine rabbits 11 days after an intravenous injection of crude papain solution was compared with that in costal cartilage from eight normal untreated rabbits. An increased fixation of the isotope was found in treated animals compared with controls.

The depletion of cartilage matrix by papain provided an experimental situation to test the hypothesis that the depletion of matrix which occurs in osteoarthritic cartilage can stimulate increased synthesis of chondroitin sulfate.

The results give further support to the view that the primary lesion in osteoarthritis occurs in the matrix rather than in the chondrocyte of articular cartilage.
PHYSICAL AND BIOLOGICAL PROPERTIES OF INFLUENZA VIRUS COMPONENTS OBTAINED AFTER ETHER TREATMENT
Authors: Davenport Fred M. , Rott Rudolf , Schäfer Werner
Published: June 2004

The Rostock strain of fowl plague, the swine, A, A', and Asian strains of influenza A as well as their hemagglutinin and internal s antigen subunits obtained after ether splitting, were found to be morphologically indistinguishable when examined simultaneously. Hemagglutinin fractions reacted in a highly strain specific manner when tested by hemagglutination inhibition or by complement fixation using sera obtained after infection. With the same sera internal s antigen fractions were shown to be serologically distinguishable by complement fixation. This observation may stimulate interest in the feasibility of employing immunologic techniques for the study of nucleoproteins. The significance of the findings reported is discussed.
PATHOGENESIS OF FEVER: EFFECTS OF VARIOUS ENDOGENOUS PYROGENS UPON THE LEVEL OF CIRCULATING GRANULOCYTES IN NORMAL RABBITS
Authors: King M. K.
Published: June 2004

The endogenous pyrogen in the serum or plasma of rabbits 2 hours after the intravenous injection of typhoid vaccine had a marked effect on the circulating leucocytes of normal rabbits. Immediately following intravenous injection there was a brief, but marked, granulocytopenia which was quickly followed by a granulocytosis. Under the same circumstances pooled heterologous serum or plasma from normal rabbits produced no fever or significant change in the level of circulating leucocytes.

The cell-free fluid of sterile peritoneal exudates produced a marked leucocytosis without a preceding leucopenia when injected intravenously into normal rabbits. When comparably pyrogenic doses of typhoid vaccine were injected in the same manner no significant change in the level of circulating leucocytes occurred.

The relevance of these findings to the pathogenesis of fever is discussed.
DISTURBED REPRODUCTIVE FUNCTION FOLLOWING SECTION OF THE PORTAL VEIN
Authors: Huggins Charles , Okamoto Ryohei
Published: June 2004

Integrity of the portal vein is a condition for normal function of the reproductive tract in female rats. Section of the portal vein (PVS) always abolished periodic sexual function. In a series of 23 animals, 6 developed continuous estrus and other signs of the presence of excessive amounts of estrogenic hormones; yet the ovaries of these rats were small. In 6 other rats there was growth of the prostate and additional evidence of increased activity of androgenic hormones. Many rats failed to become pregnant and nearly all pregnant rats aborted. Except for a decrease in lipid content, the composition of the liver of PVS rats resembled that of intact mates in the criteria which were applied.

The rapid passage of blood through the liver is necessary to conjugate the excessive amounts of steroids which the rat ovary normally produces and an intact portal vein is an essential unit in the mechanism necessary for their partial inactivation.
STUDIES OF TWO KINDS OF VIRUS PARTICLES WHICH COMPRISE INFLUENZA A2 VIRUS STRAINS
Authors: Choppin Purnell W. , Tamm Igor
Published: June 2004

Inhibitors present in normal human and animal sera prevented hemagglutination by and neutralized infectivity of inhibitor-sensitive influenza A2 virus. Starch zone electrophoresis of sera indicated that the same serum components possess both hemagglutination-inhibiting and neutralizing activities. The greatest amount of inhibitory activity was found in normal horse serum, and the inhibitory activity increased with heating or treatment with concentrated solutions of urea. The inhibitory activities on human, ferret, and rabbit sera were markedly reduced but not completely eliminated by V. cholerae filtrate and purified neuraminidase. The inhibitory activity of horse serum was only moderately reduced by these agents. The nature of the horse serum inhibitor and the differences in the interactions of inhibitor-sensitive and insensitive influenza A2 virus particles and pre-1957 influenza viruses with receptors have been discussed.
STUDIES OF TWO KINDS OF VIRUS PARTICLES WHICH COMPRISE INFLUENZA A2 VIRUS STRAINS
Authors: Choppin Purnell W. , Murphy James S. , Tamm Igor
Published: June 2004

Seven strains of influenza A2 virus were serially passed in the chick embryo, and morphological characteristics of the passages were examined in the electron microscope. With serial passage there was a change from a prominently filamentous appearance in early passages to an almost entirely spherical appearance in later passages. The number of passages required for the conversion to spherical morphology varied with different strains. The filament-sphere variation was found to be independent of the properties which differentiate "+" and "-" influenza A2 virus particles, and both highly filamentous and spherical populations of "+" and of "-" virus particles were obtained. The usefulness of these pairs of independent markers for genetic studies is discussed.
CELLULAR LOCALIZATION OF ROUS SARCOMA VIRUS AS STUDIED WITH FLUORESCENT ANTIBODY
Authors: Mellors Robert C. , Munroe J. Spencer
Published: June 2004

A study has been made of the immunospecific, apple-green staining patterns imparted by fluorescent antibody to Rous virus in frozen sections of wing tumors produced in young chickens by various initiating doses of virus. Variations in the cellular site (cytoplasm, cell membrane, and nucleus), the distribution (particulate or homogeneous), and the quantity (little or much) of Rous viral antigens were seen. The per cent of sarcoma cells containing viral antigens was related to the tumor initiating dose of virus and to the infectivity titer of virus extracted from the wing tumor. With certain assumptions it was estimated that a quantity of virus approaching one tumor-producing dose (1 TD50) per tissue section was detectable with fluorescent antibody.

Sarcoma cells containing viral antigens were identified as early as 4 days after inoculation of Rous virus. At this time viral antigens were also localized in the skeletal muscle of the inoculated wing in a characteristic focal pattern of deposition near or on the sarcolemma. This association between viral antigens and skeletal muscle fibers was observed also at 3 days after virus inoculation, when sarcoma cells were not yet demonstrable in the tissue sections.

Viral antigens were detected in sarcoma cells in lung tumors of young chickens and in tumors of the chorioallantoic membrane of chick embryos.
HOST-PARASITE FACTORS IN GROUP A STREPTOCOCCAL INFECTIONS
Authors: Cremer Natalie , Watson Dennis W.
Published: June 2004

The data presented in this study suggest that Group A streptococci produce two distinct pyrogenic toxins: one primarily an intracellular toxin and the other mainly an extracellular toxin.

Both of these toxins are not extractable by the conventional methods, phenol and TCA, used for isolation of Gram-negative bacterial endotoxin. Their chemical composition, therefore, differs from that of Gram-negative bacterial endotoxin.

On the basis of tolerance studies it appears that similar non-specific host factors may be involved in the detoxification of both the streptococcal intracellular pyrogenic toxin and the Gram-negative bacterial endotoxin. Detoxification of streptococcal exotoxin requires a more immunologically specific mechanism.

Because all the Group A sonic extracts studied elicited reciprocal tolerance to one another, it is suggested that tolerance to the intracellular pyrogenic toxin is not correlated with type-specificity.
TISSUE MAST CELLS AND ACUTE INFLAMMATION IN EXPERIMENTAL CUTANEOUS MUCORMYCOSIS OF NORMAL, 48/80-TREATED, AND DIABETIC RATS
Authors: Sheldon Walter H. , Bauer Heinz
Published: June 2004

The role of the tissue mast cells in relation to the acute inflammatory reaction to experimental cutaneous mucormycosis was studied histologically in normal rats, in animals whose tissue mast cells had been depleted of their cytoplasmic granules prior to infection by the administration of compound 48/80 and in others in whom acute alloxan diabetes with acidosis had been produced before injection of the fungus.

The discharge of the tissue mast cell granules in normal rats occurred within minutes at the site of infection and appeared to initiate the rapid onset of acute inflammation. The degranulation of the tissue mast cells subsided in a short time and the cells reassumed a normal histologic appearance while inflammation progressed with the formation of circumscribed lesions. In animals pretreated with compound 48/80 in which the tissue mast cells contained no granules, the onset of inflammation was briefly delayed, the intensity of the process was somewhat decreased, fibroblastic proliferation was retarded, and the fungus growth in the early lesions was increased. However, the infection did not spread and the lesions were well localized. The tissue mast cells in the diabetic and acidotic rats completely failed to discharge their cytoplasmic granules, the onset and intensity of the acute inflammatory response were markedly delayed and decreased and the infection progressed rapidly with massive fungus growth invading adjacent tissues. A relationship between the discharged tissue mast cell granules and eosinophilic granulocytes was noted since the latter were numerous among the inflammatory cell exudate in normal rats and scarce in the lesions of the diabetic animals.

It is concluded that a function of the tissue mast cells in the normal rat is the rapid initiation of acute inflammation at the site of injury and that degranulation of these cells prior to infection somewhat delays the inflammatory response and therefore slightly diminishes host resistance. Furthermore, a severe metabolic disorder such as acute alloxan diabetes with acidosis, inhibits the normal function of the tissue mast cells, delays and decreases inflammation, and in this manner contributes to the greatly increased susceptibility of the host to infection.
STUDIES ON IMMUNE CELLULAR INJURY
Authors: Lepow Irwin H. , Ross Ann
Published: June 2004

An initial separation of reaction stages in immune cytoxicity is described. Primary isolates of normal human amnion cells were reacted first with specific rabbit γ2-globulin antibody, then with human C'1 esterase in the presence of Ca++, and finally with human C'2, C'3, and C'4 in the presence of Mg++. Cytotoxicity, as measured by uptake of trypan blue, occurred only when these reactions were performed in the order given and did not occur when any of the constituents was omitted. Inhibition of immune cytotoxicity was achieved with partially purified preparations of human serum inhibitor of C'1 esterase under certain experimental conditions.

The data are discussed in relation to the role of C'1 esterase in initiating complement action and the partial biochemical definition of an early event in immune cytotoxicity.
STUDIES ON THE MECHANISM OF THE FORMATION OF THE PENICILLIN ANTIGEN
Authors: Levine Bernard B.
Published: June 2004

Seven highly purified degradation products of penicillin G (PG) were examined with regard to their ability to cross-react allergically with PG. Guinea pig allergic contact dermatitis was employed as the test system. Three of these degradation products, D-benzylpenicillenic acid (BPE), D-penicillamine, and D-α-benzylpenicilloic acid were found to cross-react with PG and also to be capable of inducing delayed contact allergy in the guinea pig. BPE and PG cross-reacted with particularly intense reactions, and other immunologic experiments indicated that PG and BPE introduce identical allergic determinant groups into epidermal proteins.

These experimental results were correlated with the results of previous studies concerning the degradation pathways of PG under physiological conditions in vitro, and the chemical reactivities of these degradation products.

Based on these immunologic and chemical data, a schema is proposed which suggests the chemical pathways by which PG may react with epidermal proteins in vivo to form the penicillin antigen. The identity of the specific antigenic determinant groups of the penicillin antigen is suggested.

The relationship between PG allergy of the contact dermatitis type in the guinea pig and PG allergy of the immediate type in man is discussed.
THE LEVEL OF LACTIC DEHYDROGENASE ACTIVITY AS AN INDICATOR OF THE GROWTH OF INFLUENZA VIRUS IN THE EMBRYONATE EGG
Authors: Kelly Richard , Greiff Donald
Published: June 2004

The relation, between time and the levels of lactic dehydrogenase activity of chorioallantoic fluids from embryonate eggs infected with the PR8 strain of influenza virus were determined quantitatively. The mean values, based on 10 determinations for each time interval, followed a sigmoid curve, with the greatest rates of change occurring between 48 and 72 hours after the inoculation of virus. The activities of the fluids from infected eggs at the 72nd hour or later were approximately 18 times higher than those from non-infected eggs. Based on the data above, a qualitative test for the presence of infection with influenza virus was developed.
PREPARATION OF AN INHIBITOR OF VIRAL HEMAGGLUTINATION FROM HUMAN ERYTHROCYTES
Authors: Kathan Ralph H. , Winzler Richard J. , Johnson Clarence A.
Published: June 2004

A material, derived from human erythrocytes and believed to be identical with the receptor site for the myxoviruses, has been obtained in homogeneous form. The method of preparation involves pH adjustment of the stroma, hot phenolic extraction, chloroform-methanol treatment, and ultracentrifugation. The material so obtained possesses a high inhibitory titer to viral hemagglutination (45,000 to 60,000 inhibitory units/mg. against 4 hemagglutination units of Lee strain of influenza virus) and appears to be a glycoprotein containing 22 to 24 per cent sialic acid, 12 per cent hexose, 12 per cent hexosamine, 1 per cent fucose in addition to at least eleven amino acids. The sialic acid probably occupies a terminal position in an oligosaccharide chain extending from the protein peptide chain. The molecular weight is near 30,000—an unusually low value for substances possessing this biological activity. M and N blood group activity also seems to be associated with this protein.
GLOMERULAR PERMEABILITY
Authors: Farquhar Marilyn G. , Wissig Steven L. , Palade George E.
Published: June 2004

Ferritin was used as a tracer to investigate pathways and mechanisms for transfer across the various layers of the glomerular capillary wall. Kidney tissue, fixed at intervals of 2 minutes to 2 hours following an intravenous injection of ferritin, was examined by electron microscopy.

The observations confirmed the existence of three distinct and successive layers in the glomerular capillary wall (the endothelium, the basement membrane, and the visceral epithelium). In addition, they demonstrated a number of new structural features: namely (a) discrete fibrils in the subendothelial spaces; (b) a characteristic, highly elaborate, cytoplasmic organization in the visceral epithelium; and (c) special structures resembling "desmosomes" in the slits between foot processes.

In animals sacrificed at short time intervals (2 to 15 minutes) following ferritin administration, ferritin molecules were found at high concentration in the lumen and endothelial fenestrae, at low concentration in the basement membrane, and in very small numbers within the epithelium.

Later (1 to 2 hours), the tracer particles were still present in the lumen and within endothelial fenestrae, and, in addition, had accumulated on the luminal side of the basement membrane, especially in the axial regions of the vessels. Larger numbers of ferritin molecules were also found in the epithelium—in invaginations of the cell membrane at the base of the foot processes, and in various membrane-limited bodies (vesicles, multivesicular bodies, vacuoles, and dense bodies) present within the cytoplasm.

These observations suggest that the endothelial fenestrae are patent and that the basement membrane is the main filtration barrier. Since the basement membrane has no demonstrable pores, it is probably not a simple sieve but presumably is a gel-like structure with two fine fibrillar components embedded in an amorphous matrix. Both the epithelium and endothelium may be concerned with building and maintaining this structure. Finally, the intracellular accumulation of particles in the epithelium suggests that the latter acts as a monitor that recovers, at least in part, the small amounts of protein which normally leak through the filter.
NUCLEIC ACID SYNTHESIS IN TYPES 4 AND 5 ADENOVIRUS-INFECTED HELA CELLS
Authors: Ginsberg Harold S. , Dixon Mary K.
Published: June 2004

Type 4 adenovirus infection of HeLa cells effected a marked increase in synthesis of the saline-soluble DNA fraction, but not the host-cell DNA (the water-soluble fraction). This was demonstrated by the marked increase in specific activity of saline-soluble DNA but not water-soluble DNA when P32-inorganic phosphate or sodium formate-C14 was employed. When these isotopes were used to label cells before viral infection rather than during the process of viral propagation, the saline-soluble DNA from infected cells had a specific activity of 10 to 20 per cent less than that of uninfected cells, indicating that the saline-soluble DNA was synthesized both from prelabeled precursors of the cell pools and unlabeled materials from the medium. Saline-soluble DNA began to increase between 10 to 12 hours after viral infection and 3 to 4 hours before appearance of newly propagated infectious virus. The specific activity of the acid-soluble pool of infected cells also increased between 10 to 12 hours after viral inoculation when sodium formate-C14 was used as a radioisotope. When P32-inorganic phosphate was utilized, the specific activity of infected-cell RNA was increased approximately the same relative amount as when total RNA was determined chemically; i.e., 30 to 40 per cent. With type 5 adenovirus, not only did a 3- to 5-fold increase in saline-soluble DNA occur, but also an increase was measured in specific activity of RNA when P32-inorganic phosphate was used.
EFFECT OF MULTIPLICITY OF INFECTION ON NEWCASTLE DISEASE VIRUS-HELA CELL INTERACTION
Authors: Wheelock E. Frederick , Tamm Igor
Published: June 2004

The effects of a hundred-fold difference in virus/cell multiplicity on the interaction of Newcastle disease virus (NDV) with HeLa cells were studied, and various phases of the virus reproductive cycle were related to cellular consequences of infection. At both multiplicities used all cells were infected. The following events occurred 1 to 2 hours earlier in cells which were inoculated with the higher multiplicity: (a) first appearance of newly made virus antigen, and the amount present at any time during the period of rapid increase; (b) onset and time course of production of infective virus; (c) development by infected cells of hemadsorbing ability; (d) onset and time course of inhibition of mitosis; and (e) onset and time course of marked cell damage.

Double infection of HeLa cells with NDV and NWS was demonstrated by the fluorescent antibody technique, and was used to show that the establishment of interference against NWS was also dependent upon the multiplicity of NDV.

In cells inoculated at each multiplicity, newly made virus antigen appeared at the same time as the first infective virus particles. Infective virus rapidly reached a peak, and then declined. Viral antigen continued to increase for several hours after the decline in infective virus had begun. Thus, only a small fraction of the virus antigen produced was incorporated into new infective particles. The maximal yield of such particles was only 6 to 11 per HeLa cell. Over 95 per cent of new virus was cell-associated, but could be neutralized by treatment with antiserum before disruption of cells.

Mitosis occurred in cells which had produced and released infective NDV. Progressive inhibition of mitotic activity in infected cells was correlated with continued production of viral antigen. Marked cytopathic changes developed after mitotic activity had decreased to low levels.

The mechanism by which NDV inhibits mitosis in HeLa cells is discussed.
A COMPARATIVE STUDY OF THE GROWTH, NUTRITION, AND METABOLISM OF THE PRIMARY AND THE TRANSFORMED HUMAN CELLS IN VITRO
Authors: Chang R. Shihman
Published: June 2004

The growth characteristics, nutritional requirements, and metabolic activities of primary explants (chiefly amnion) and transformed human cells (derived from amnion, conjunctiva, and cervical cancer) were compared. Major differences observed are as follows:

1. Five types of human tissue when freshly explanted in vitro were found to have varying but limited growth potential in contrast to the autonomous and apparently unlimited growth of the transformed cell.

2. A requirement for an external source of inositol or glutamine could not be demonstrated for the slowly multiplying primary amnions in contrast to the rapidly growing transformed cells which degenerated in the absence of either metabolite.

3. Unlike the transformed cells, the primary amnion cells fixed an insignificant amount of CO2; when infected by the Coxsackie virus, however, a requirement for CO2 in the formation of more virus became readily demonstrable.

4. The biggest difference in the incorporation of various C14 substrates occurred in the nucleic acid fractions.

5. The difference in the incorporation of glycine into the protein fraction was very small.

6. The utilization of glucose, ribose, and xylose as well as the oxidation of lactic acid into CO2 by the primary amnion cells occurred at a rate much lower than that of the transformed cells.

7. The cytotoxic dose of 5-fluorodeoxyuridine for the primary amnion was at least 100 times that of the transformed cells although growth inhibition was readily demonstrated at 0.1 µg. per ml. for both types of cells.

Based on these observations, several hypotheses are put forth for consideration.
A SIMPLIFIED CHEMOSTAT FOR THE GROWTH OF MAMMALIAN CELLS: CHARACTERISTICS OF CELL GROWTH IN CONTINUOUS CULTURE
Authors: Cohen Edward P. , Eagle Harry
Published: June 2004

A simplified technique has been described for the continuous growth of mammalian cells in suspension culture. The cell population density increased as the rate of input of fresh medium was decreased, and the average generation time was concommittantly prolonged. At relatively high input rates, the population remained stabilized for an indefinite period, but at low flow rates, there was sometimes a cyclical variation in population density. The factor limiting growth rate at input rates of approximately 0.2 volumes per day was not the exhaustion of the medium; but in some experiments a non-dialyzable material appeared which inhibited cell growth.
RHEUMATOID FACTOR AND THE PATHOGENESIS OF RHEUMATOID ARTHRITIS
Authors: Mellors Robert C. , Nowoslawski Adam , Korngold Leonhard , Sengson Beth L.
Published: June 2004

In analogy with the two categories of reactants which are used in the serological tests for the unusual category of macroglobulins called rheumatoid factor, two fluorescent reactants have been prepared for the detection of rheumatoid factor in situ in tissue sections: fluorescent antigen-rabbit antibody (immune) complex, in the present study, and fluorescent aggregated human γ-globulin, in previous work.

Plasma cells in the synovial membrane and germinal center cells and internodular plasma cells in lymph nodes are the sites of origin of rheumatoid factor in active rheumatoid arthritis, whether occurring in adults or children. Plasma cells and germinal center cells which form rheumatoid factor detectable with fluorescent immune complex are less numerous than those which contain factor demonstrable with fluorescent aggregate. In the same tissues, plasma cells and germinal center cells which contain macroglobulin (19S human γ-globulin) detectable with fluorescent antibody—but not showing the reactivity of rheumatoid factor—are more abundant than those containing rheumatoid factor.

While macroglobulin and rheumatoid factor are almost exclusively formed in the cytoplasm, these proteins are also detectable in the nucleus of an occasional plasma cell.

Normal and pathological synovial and capsular tissues, lymph nodes, and connective tissues obtained from individuals without rheumatoid arthritis are not stained with fluorescent immune complex or, except for an unusual example of Waldenstrom's macroglobulinemia, with fluorescent aggregate.

The cellular origin, as well as certain chemical and immunological attributes, of rheumatoid factor suggests an antibody-like nature and function. The observations cited are consistent with the behavior anticipated for cellular rheumatoid factor, were it primarily an antibody direct to an altered human γ-globulin and cross-reacting with rabbit γ-globulin. However, it is also possible that there are two or more cellular rheumatoid factors.

Lesion-associated protein precipitates having the composition anticipated for rheumatoid factor-antigen complex are localized in the amyloid depositions in kidney and spleen of an individual who died with amyloidosis secondary to rheumatoid arthritis.
STUDIES OF ARTHRITIS AND OTHER LESIONS INDUCED IN RATS BY INJECTION OF MYCOBACTERIAL ADJUVANT
Authors: Pearson Carl M. , Waksman Byron H. , Sharp John T.
Published: June 2004

A generalized disease is induced experimentally in the rat by administration of Freund's adjuvant. The primary clinical and pathologic lesions are arthritis, periarthritis, peritendinitis, and periostitis in the joints of the extremities and tail. Accompanying the arthritis in some cases, and never observed in its absence, are other specific tissue lesions including iridocyclitis, nodular lesions in the glabrous skin (ear, genitalia, feet, tail), transient rashes, a chronic skin disease, genitourinary lesions, and diarrhea. These make up a striking and characteristic picture. The arthritis usually precedes the other lesions and, together with the skin disease may show a prolonged and fluctuating course. Visceral lesions do not occur.

Histologically, the basic lesion is a lymphocytic and histiocytic infiltration, initially perivascular and subsequently more disseminated. In addition, in the region of the joints and in the corpora cavernosa of the penis, there is extensive proliferation of mesenchymal cells, especially fibroblasts. Foci of fibrinoid necrosis are seen in the articular and nodular lesions, and destructive lesions of the joints are common.

Such a combination of tissue lesions has not previously been described in experimental pathology. The experimental disease is shown to have both similarities to and differences from Reiter's syndrome, rheumatoid arthritis, and certain other disorders which occur in man.
THE INTERACTION OF MAMMALIAN CELLS WITH ANTIBODIES. I
Authors: Oda M. , Puck T. T.
Published: June 2004

The single cell plating technique has been applied to quantitation of the reproductive killing of mammalian cells by specific antibodies. This method confirms previous demonstrations by other workers of localization of all the killing activity in the γ-globulin fraction of specific cell antisera but not of normal sera; the need for complement for the killing action in low doses of antibody and the leakage of cell constituents attending cell killing under these conditions. In concentrations of 4 per cent or higher of heated antiserum cell killing occurs without added complement. The cell plating technique permits highly reproducible quantitation of antibody action and demonstrates antibody activity in sera diluted 1:3000. It permits demonstration of very high degrees of species specificity as shown by virtually complete absence of cross-reaction between antisera to Chinese hamster and S3 HeLa cells, respectively. Somatic cells which have been sensitized by absorption of specific antibody lose their sensitization when incubated at 37° unless complement is added within 1 hour.
CELLULAR SOURCE AND CHARACTER OF A HEAT-STABLE BACTERICIDAL PROPERTY ASSOCIATED WITH RABBIT AND RAT PLATELETS
Authors: Jago Richard , Jacox Ralph F.
Published: June 2004

Rabbit and rat platelets contain two non-dialyzable components (analogous to Myrvik's components 1 and 2 of serum) which, together with bicarbonate ions, produce a potent bactericidin for B. subtilis. These platelet factors appear to be identical with the serum bactericidal substances, suggesting that disruption of platelets during blood coagulation releases the platelet factors into serum. Normal human platelets contain no measurable component 1 or 2.

Since bovine serum contains component 2 but little or no component 1, estimation of the concentration of component 1 in sera or platelet suspensions of other species can be made by diluting the materials to be tested in bovine serum. The level at which a bactericidal effect is lost represents the end-point of the titration of component 1.

Rabbit serum has a high concentration of component 1 and a lower concentration of component 2. It can be diluted to a point at which bactericidal activity is lost, owing to a depletion of component 2, but at which component 1 is still present in effective concentration. At this point the addition of another serum or a platelet homogenate causes a restoration of the bactericidal activity and permits an estimate of the concentration of component 2 in the added material.