Ultrastructure of lipopolysaccharide isolated from Thermoplasma acidophilum. The fine structure of lipopolysaccharide isolated from Thermoplasma acidophilum was examined by electron microscopy. Negative staining of the lipopolysaccharide revealed long, ribbon-like structures with some branching. The average width of the lipopolysaccharide ribbons was 5 nm. Treatment of the lipopolysaccharide with 0.5% sodium dodecyl sulfate resulted in the dissociation of the ribbon-like structures to spherical- and vesicular-shaped particles and some short, rodlike structures. Results suggest that the lipopolysaccharide from T. acidophilum is morphologically similar to lipopolysaccharide isolated from gram-negative bacteria.


The surgical correction of Fallot's tetralogy. The success of total repair of tetralogy of Fallot depends on a clear understanding of the morbid anatomy and deranged hemodynamics of this anomaly, which consists essentially of two abnormalities, namely, a large unrestrictive ventricular septal defect and a severe obstruction to the outflow of the right ventricle. The ventricular septal defect acts as a safety valve and, therefore, cannot be closed unless the obstruction to the outflow of the right ventricle is adequately relieved. In severe cases, this often necessitates reconstruction of the outflow. With an adequate bypass technique and a clear understanding of the principles described in this article, complete correction of tetralogy of Fallot can be accomplished with a low mortality. In a series of 100 consecutive patients presented, the mortality was only 3%.


Long-term results with the Björk-Shiley tilting disk valve in aortic valvular disease. The construction of the Björk-Shiley prosthesis, with its nonoverlapping tilting disk, provides an optimal orifice area in relation to tissue diameter. The transprosthetic forward flow is central and mainly laminar. The resistance to blood flow remains low with increasing cardiac output during exercise, even when the smaller sizes of the prosthesis are used for implantation in narrow aortic roots. Mechanical crushing of erythrocytes is minimized by the nonoverlapping closing mechanism, and the resulting regurgitation is negligible. The in vivo durability of the prosthesis is excellent and thromboembolism does not occur as long as dicoumarol treatment is effective. In aortic valvular disease, an effective unloading  of the left ventricle was obtained at rest and during exercise after aortic valve replacement. The resulting hemodynamic improvement correlated with a reduction in cardiomegaly, increase in physical working capacity and relief of distressing symptoms. Clinical improvement was maintained for up to five years.


Transposition with ventricular septal defect and pulmonary stenosis: the Rastelli operation. A total of 52 Rastelli operations have been performed at the Mayo Clinic for transposition of the great arteries with ventricular septal defect and pulmonary stenosis. Of the 43 patients over four years of age, two died (5%). A much higher mortality among younger children, in our earlier experience, led to a policy of early palliation, if required, by a Blalock-Taussig shunt, with later correction. The technique which evolved is described. The results of the operation thus far suggest that its success is comparable to that for the repair of the tetralogy.


Energy coupling in the uptake of hexose phosphates by Escherichia coli. Several methods were used to study the source of energy in the uptake of hexose phosphates by Escherichia coli K12. The uptake was sensitive to inhibition by agents that affect electron transport, such as lack of oxygen, cyanide, and heptylhydroxyquinoline-N-oxide, and by agents that affect ATP utilization, such as dicyclohexylcarbodiimide and arsenate. It was also sensitive to uncouplers in the presence of absence of oxygen. The strain of E. coli used extruded protons during respiration. Uncer anaerobic conditions, the uptake of approximately 1 eg to H+ per glucose 6-phosphate. These observations are consistent with a chemiosmotic mechanism of genergized glucose 6-phosphate uptake. The rate of glucose 6-phosphate uptake was maximal in KC1, but was also stimulated by MgC12 or CaC12. Inhibition by A217, a nigericin-like antibiotic, was prevented by K+ whereas valinomycin and gramicidin inhibited in the presence or absence of K+.


Serological analysis of human deoxyribonucleic acid polymerases. Preparation and properties of antiserum to deoxyribonucleic acid polymerase I from human lymphoid cells. The preparation and properties of an antiserum to human DNA polymerase I (6 to 8 S) are described. Care was taken in the purification of the antigen to remove certain other DNA polymerases found in human cells. An incubation of antigen and antiserum lasting about 48 hours is necessary to achieve maximal inhibition. About 1 mug of the antipolymerase immunoglobulin G, prepared in rats, neutralizes 60% of the activity present in 54 ng of the enzyme. Tritrations varying both antiserum and enzyme demonstrate clear regions of antigen and antibody excess. Inhibition of enzyme activity is about the same whether the templateprimer is (dA)n-(dT)12-18, or partially digested DNA. An assay was developed which measures the remaining activity in the supernatant after precipitation of enzyme-antibody complexes with goat anti-rat immunoglobulin G. In this assay, 2.2 mug of the antipolymerase immunoglobulin G quantitatively bind 33 ng of DNA polymerase I. With use of the direct neutralization assay and the immuno-precipitation test, we found little, if any, antigenic relationship between DNA polymerase I and DNA polymerase II (3.4 S). Similarly, little, if any, relationship was found to the DNA polymerases from five RNA tumor viruses. The activities of RNA-directed DNA polymerases from the blood leukocytes of two patients with acute myelogenous leukemia and from the placentas of rhesus monkeys were not inhibited in neutralization assays which were shortened because these enzymes were thermolabile. In identically shortened neutralization assays, the antipolymerase immunoglobulin G neutralized up to 76% of the activity of DNA polymerase I. In addition to its utility in distinguishing cellular DNA polymerases, the rat antiserum should be useful reagent for testing of novel DNA polymerases isolated in small quantities from human tumors for contamination with DNA polymerase I. This enzyme is present in abundance in proliferating tissue and often confuses the biochemical characterization of these novel enzymes.


Allergic encephalomyelitis. Isolation of an encephalitogenic peptide active in the monkey. A 17-residue peptide (Peptide Y) was isolated from the COOH-terminal end of the basic protein of bovine myelin by peptic digestion. This peptide induced experimental allergic encephalomyelitis in the rhesus monkey. Treatment of Peptide Y with cyanogen bromide released three amino acids from the COOH-terminal end and resulted in a tetradecapeptide (Peptide M) which was also encephalitogenic in the rhesus monkey. The sequence of Peptide M is: Phe-Lys-LEU-Gly-Gly-Arg-Asp-Ser-Arg-Ser-Gly-Ser-Pro-Met. Thus a major disease-inducing site active in the rhesus monkey is contained within a 14-residue peptide localized near the COOH-terminal end of the protein. This peptide differs markedly in location and sequence from the 9-residue peptide shown to contain the encephalitogenic determinant for the guinea pig.


The reaction of mitochondria in the coleoptiles of rice (Oryza sativa L.) with diaminobenzidine. Diaminobenzidine, DAB, was applied to segments of aerobically and anaerobically grown coleoptiles of rice, Oryza sativa L., with the object of studying the location of cytochrome oxidase at the electron-microscope level. A specific staining of mitochondrial cristae and inner membrane was obtained, with no reaction in other organelles; with extended periods of incubation, the reaction product filled the mitochondria completely. In anaerobically grown coleoptiles, the reaction was much slower and the difference was particularly marked in vascular bundle companion cells and parenchyma, which gave the strongest reaction in aerobic tissue, but in the anaerobic stained even less than the cortical parenchyma. The reaction was inhibited by boiling and slowed very much by lowering of the incubation temperature from 27 to 4 degrees C. This indicated the involvement of an enzymic reaction and cyanide inhibition indicated that a haem enzyme was involved. The catalase inhibitor aminotriazole did not inhibit DAB oxidation. Nevertheless the specificity of the reaction for cytochrome oxidase must be questioned, because preheating of the tissue to 60 degrees C before incubation, which would be expected to destroy cytochrome oxidase activity, failed to decrease the oxidation, at least in aerobically grown coleoptiles. It is concluded that DAB is oxidized in the rice coleoptile tissue by a cytochrome system, and the development of this system is inhibited by anaerobiosis, but the oxidation cannot be claimed to represent cytochrome oxidase activity exclusively. Perhaps other autoxidizable, more heat-stable cytochromes participate in the reaction.


Reconstruction of axonal trajectory of individual neurons in the spinal cord using Golgi-stained serial sections. A procedure bringing the axonal and dendritic cut ends found in the confronting planes of the serial sections face to face by means of serial photomicrographs provides a means for precisely reconstructing individual Golgiimpregnated neurons. Utilizing this method, it was possible to follow at the caudal level of a kitten's spinal cord the total course of the axons and dendrites of sixteen intramedullary neurons. The neurons located in the dorsal horn (laminae IV and V) send their long ascending axon into the lateral funiculus of the ipsi- or contralateral side, and, in one case, into the anterior funiculus of the ipsilateral side, giving off on the way several collaterals to the dorsal half of lamina VII of the ipsi- or contralateral side. Some of the neurons found in the intermediate region and the ventral horn (laminae VII, VIII and X) distribute their axonal branches in the intermediate region or the ventral horn of the ipsi- or contralateral side or both sides, running throughout their course in the grey matter, while the others act in the same manner by way of the lateral or anterior funiculi. Axonal and collateral endings terminate with small terminal knobs. The total axonal length of the neurons observed, with the exception of some with a long ascending trajectory, varies from 1,680 mum to 6,480 mum.


The distribution of the alpha type of ganglion cells in the cat's retina. There is a distinct class of large ganglion cell bodies observable in whole-mount preparations of the cat's retina stained with cresyl violet. Measurements of perikaryal size and its variation with eccentricity from the central area support the identification of the large cells with the class of alpha cells previously defined in Golgi-stained preparations. A complete enumeration of alpha cells in one retina yielded a total of 6212. They were encountered in all parts of the retina and had a peak density within the central area (  200/mm2). The retinal distribution was displayed as a contour map of alpha-cell density. Away from the central area, the lines of iso-density had the form of a 4-pointed star with rather blunt points corresponding to horizontal and vertical ridges of augmented density. Along horizontal, vertical and oblique strips through the central area, alpha-cell density was an approximately constant fraction of total ganglion cell-density. The average value of the fraction was 3.3% from which it has been calculated that the cat's retina may contain as many as 190,000 ganglion cells.


On the "selectivity" of the Golgi-Cox method. The randomness of the impregnation of layer IV cortical neurons by the Golgi-Cox method (Van der Loos, '56) has been assessed directly in Barrel C-1 of the mouse SmI. All Golgi-Cox impregnated neurons and unimpregnated neurons which were revealed with a Nissl counterstain were counted and measured in ten cerebral hemispheres cut tangential to the pia overlying the barrel field. The percentage of Golgi stained neurons varied considerably in different preparations from 0.73% to 2.26% with an average of 1.29%. The size distributions of both the Golgi impregnated and Nissl stained cells are similar but the difference of the means is statistically significant. However, if the means are equated there is no statistical difference in the two populations. When the Golgi precipitate is removed and the cells re-measured following Nissl staining there is a systematic reduction of the perikaryal cross-sectional area which is compatible with the differences in the means observed for the two populations as a whole. Finally, the frequency with which Golgi impregnated neurons are found in the barrel sides and hollows parallels the frequency with which Nissl stained neurons are observed in these two locations. We conclude that this variant of the Golgi method impregnates barrel neurons randomly. The value of this information for quantitative studies of cerebral cortex is discussed as is the potential of the system for elucidating some of the mechanisms responsible for Golgi impregnation.


Subtypes of hepatitis B antigen among patients and symptomless carriers. A study of the distribution of subtypes ad and ay among sera from hepatitis B antigen-positive subjects in North West England and North Wales revealed a marked contrast between symptomless carriers among whom ad predominated and patients with acute hepatitis the majority of whom were ay. Those with hepatitis associated with drug addiction or other forms of "needle transmission" were almost all of subtype ay. On the other hand in cases of "sporadic" hepatitis without evidence of parenteral exposure subtypes ad and ay are about equally distributed. These findings are similar to those reported from other countries in Northern Europe and North America. Although geographical and social factors clearly affect the distribution of the two subtypes it is suggested that the virus of subtype ay may be more readily transmitted than subtype ad by parenteral routes involving small amounts of blood.


Catabolism physical, and immunologic properties of endocytosed isologous and heterologous iota-globulins by mouse macrophages. The catabolism of completely endocytosed isologous and heterologous gamma-globulins by mouse macrophages was studied in vitro. Mouse, human, and rabbit 125-I-IgG were coupled to mouse, human, or sheep erythrocytes either as antibodies or by covalent binding. They were exposed to macrophages for 1 hr and the non-endocytosed erythrocytes were then removed with a Ficoll gradient centrifugation. Catabolism was evaluated after 2, 5, and 18 hr in culture by measuring the radioactivity released into the culture medium as well as the radioactivity that remained associated with cells. It was found that all iota-globulins were catabolized in a similar fashion, and that the type of carrier erythrocytes (isologous or heterologous) had no influence on catabolism. Some of the material that remained associated with macrophages was on the cell membrane and could be removed by trypsin. Some of the material that was released by macrophages was completely degraded but some was either not degraded or only partially degraded. Sucrose density gradient analysis and SDS-polyacrylamide gel electrophoresis showed that this material had kept some physical properties of native iota-globulins. It was also found with the antigen-binding inhibition test and incubation with erythrocytes that the released material contained molecules carrying Fab determinants and was able to bind specifically to erythrocytic antigens. Taken together, these observations show that iota-globulins phagocytosed in the form of antigen-antibody complexes are only incompletely degraded and that the material associated with plasma membrane of macrophages or found in the culture medium is a product of cell catabolism.


Carrier-determined tolerance in vitro.

Carrier-determined tolerance in vitro. A primary immune response to normal BDF1 spleen cells was obtained in vitro to the T-independent antigen TNP-T4 coliphage. This anti-TNP response was suppressed by exposing the spleen cells for 6 hr to TNP bound either to isologous or heterologous gamma-globulin. The suppression was hapten specific. In contrast, TNP-albumin conjugates did not induce tolerance in vitro.


Spontaneous and methylcholanthrene-enhanced thyroiditis in BUF rats. II. Induction of experimental autoimmune thyroiditis without completed Freund's adjuvant. The immunization of Buffalo strain (BUF) rats with rat thyroid extract and Bordetella pertussis vaccine without complete Freund's adjuvant resulted in experimental autoimmune thyroiditis (EAT) in 90% of the rats by the 10th day after immunization. The treatment did not elicit thyroiditis in Lewis strain (LEW) rats. This susceptibility to the induction of EAT without the use of complete Freund's adjuvant was inherited by F1 hybrids of BUF and LEW. Neither histamine sensitizing factor from B. pertussus nor Escherichia coli vaccine substitute for whole pertussis vaccine.


Antigenic relationship between the fourth component of human and guinea pig complement. Antigenic similarity between cell-bound human and guinea pig C4 was detected with antisera specific for each of these molecules. No cross-reaction could be detected between C4 of normal guinea pig or human sera or after treatment of the sera with antigen-antibody C1 complexes. The cross-reactivity between cell-bound human and guinea pig C4 may reflect conformational changes in the C4 molecule as a result of binding of the cell surface.


Dissociation of the immunologic release of histamine and slow reacting substance of anaphylaxis from human lung using cytochalasins A and B-1. Cytochalasins A and B at low doses resulted in enhancement of the antigen-induced and anti-IgE-mediated release of histamine from human lung fragments in vitro. Both compounds appeared to effect a dose-dependent inhibition of the antigen-induced release of slow reacting substance of anaphylaxis. Although the effects observed with cytochalasin B were at least partially reversible, the effects of cytochalasin A could not be readily reversed. The cytochalasins appeared to be more effective when present during the antigen-dependent activation phase of mediator release rather than during the calcium-dependent release phase.


Antigen-induced inhibition of experimental allergic encephalomyelitis. III. Localization of an inhibitory site distinct from the major encephalitogenic determinant of myelin basic protein. Defined peptide fragments were isolated from the N-terminal half of the myelin basic protein (BP) molecule and employed for antigen-induced inhibition of experimental allergic encephalomyelitis (EAE). Guinea pigs pretreated with peptide 44-89, obtained by limited pepsin digestion and purified by column chromatography, were significantly protected against EAE subsequently induced by sensitization with BP in complete Fruend's adjuvant. Peptide 1-20, derived by cyanogen bromide cleavage, did not inhibit EAE, nor did the synthetic EAE peptide (residues 114-122), although this peptide was only weakly encephalitogenic for guinea pigs. These findings directly support our previous conclusion that different sites on the BP molecule are responsible for induction and inhibition of EAE, and suggest that disease inhibition can be attributed, at least in part, to a site within peptide 44-89.


Characterization of cytotoxic effector cells in the mouse mammary tumor system. Cell types involved in the immune response in vitro to MTV-induced BALB/cfC3H mammary tumors have been studied with techniques designed to inactivate or deplete different cytotoxic effector cells from spleen cells populations. The minimal activity of spleen cells from neonatally MTV-infected virgin BALB/cfC3H females is dependent upon the presence of T cells. Spleens from multiparous BALB/cfC3H females bearing small tumors are similar to those of tumor-free multiparous females. However, after the tumors grow to large size, the activity of the non-T cell population is no longer detectable. The cytotoxic activity of spleen cells from BALB/c females older than 14 weeks (supposedly not infected with MTV) is not dependent upon the presence of T cells. No activity is shown by spleen cells from BALB/c females younger than 14 weeks. Positive control spleen cells were obtained from C3H female mice who had been immunized to BALB/c histocompatibility antigens by a skin allograft; both a T cell and a non-T cell response were found.


Immunologic and biochemical properties of the histamine-sensitizing factor from Bordetella pertussis. A highly potent extract of the histamine sensitizing factor (HSF) of Bordetella pertussis was isolated by extraction of bacterial cells with urea buffer and subsequent gel filtration. This preparation of HSF also contained leukocytosis-promoting activity and adjuvant activity for reaginic and hemagglutinating antibodiesl Digestion of this extract with pronase or trypsin partially destroyed histamine-sensitizing activity, leukocytosis-promoting activity, and adjuvant activity for reaginic antibody, but did not affect adjuvant activity for hemagglutinating antibody. Antisera to HSF was prepared by immunizing rabbits with either whole bacteria or partially purfied extract. These antisera contained several precipitating antibodies to Bordetella pertussis extract demonstrated by immunodiffusion and immunoelectrophoresis. Antisera added in vitro to Bordetella pertussis extracts or passively administered in vivo to mice, reduced or abolished all biologic activities except adjuvant activity for hemagglutinating antibody. These results suggest that HSF might be an antigenic component of Bordetella pertussis which also possesses leukocytosis-promoting activity and adjuvant activity for reaginic antibody.


Characterization of guinea pig mitogenic factor. Guinea pig mitogenic factor (M.F.) was identified in antigen-stimulated guinea pig lymph node lymphocyte cultures by the ability of supernatants to stimulate DNA synthesis in non-immune lymphocytes. Its activity is not augmented by the presence of antigen and it will stimulate lymphocytes made unresponsive to the eliciting antigen. M.F. production by lymphocytes is detectable within 6 hr after antigen stimulation and reaches a maximum at 24 hr. DNA synthesis produced by M.F. is maximal at 48 hr. M.F. is heat stable at 60 degrees C. Its molecular weight is in the range of 20,000 to 30,000 daltons. M.F. activity appears to be relatively resistant to the action of proteolytic enzymes.


Specific isolation and characterization of antibody directed to binding site antigenic determinants. The preparation and specificity of antibodies specific for the ligand-binding site of HOPC 8, a phosphorylcholine (PC)-binding mouse myeloma protein, are described. Antiserum to HOPC 8, prepared in rabbits, was adsorbed with an HOPC 8-Sepharose immunoadsorbent and anti-binding site antibodies were eluted with PC. These antibodies reacted with HOPC 8 but not other myeloma proteins, including those with PC-binding specificity different from HOPC 8; the specificity of this anti-HOPC 8 antibody for the combining site region of HOPC 8 was shown by the fact that 1) the interaction of the anti-HOPC 8 antibody preparation with HOPC 8 was completely blocked by PC and 2) the antibody preparation failed to bind TEPC 15 in which the combining sites had been blocked by covalently bound PC groups. Moreover, these anti-binding site antibodies did not react with isolated heavy or light chains, indicating the requirement for a heavy-light chain interaction. By contrast an idiotypic antiserum to HOPC 8 prepared in A/J mice did bind affinity-labeled TEPC 15 and the reaction with HOPC 8 was only marginally hapten inhibitable. Both of the idiotypic determinants detected by these two antisera were present on anti-PC antibody raised in BALB/c mice;


The release of four mediators of immediate hypersensitivity from human leukemic basophils. The availability of a patient with basophilic leukemia manifesting 75 to 90% mature basophils permitted the use of a cell concentration sufficient to generate and release mediators upon interaction with a calcium ionophore in quantities adequate for their physiocochemical characterization. The mediators were defined in terms of their physicochemical characteristics: slow reacting substance of anaphylaxis (SRS-A) by purification through silicic acid chromatography and inactivation by arylsulfatase; eosinophil chemotactic factor of anaphylaxis (ECF-A) by its gel filtration through Sephadex G-25 and inactivation by subtilisin and not trypsin; and platelet-activating factor (PAF) by its inherent binding to albumin. Both ECF-A and histamine were present in their preformed state, and for histamine it was possible to establish that the concentration per cell was comparable to that of normal human basophils. Dibutyryl cyclic AMP suppressed release of histamine and SRS-A, indicating that their availability was under a control similar to that observed with normal cells subjected to immunologic activation. The demonstration that a suspension of leukemic human basophils contained the preformed mediators, histamine and ECF-A, and generated SRS-A and PAF for release along with histamine and ECF-A, after activation with a calcium ionophore, establishes that a single cell type can serve as a source of the four recognized mediators of immediate-type hypersensitivity.


Comparison of the ocular effects of circulating endotoxin and immune complexes: role of vasoactive amines. Both bacterial endotoxin and soluble immune complexes (BGG-anti BGG) injected i.v. in rabbits produce an alteration in ocular vascular permeability confined primarily to the vessels of the iridial portion of the ciliary processes. These effects have been measured by the ocular accumulation of 125I-albumin relative to cardiac plasma. Ten micrograms per kilogram of Escherichia coli bacterial endotoxin (055:B5) produce a consistent alteration in ocular vascular permeability over a 90-min period after injection. Fifty to 100 mmug/kg of endotoxin result in a prolonged effect that is maximum 4 hr after injection. Large quantities of immune complexes (BGG-antiBGG) prepared in 20 to 25 times antigen excess produce an anaphylaxis and approximately a 70% reduction in platelets and CH50. The alteration in ocular vascular permeability is half as great as that produced by 10 ug/kg of endotoxin over the 90-min period after their injection. A combined treatment with antihistamine, pyrilamine maleate, and antiserotonin, methysergide maleate, results in a significant reduction in ocular 125I-albumin in normal animals, virtually prevents the ocular effect of immune complexes, and reduces the average effect of endotoxin by 30%. Increasing the quantities of injected antihistamine and antiserotonin does not have a further effect on the response to 10 ug of endotoxin.


Immunologic properties of bacterial lipopolysaccharide (LPS): correlation between the mitogenic, adjuvant, and immunogenic activities. Bacterial lipopolysaccharide (LPS) was demonstrated to have the capacity in mice to enhance the response to soluble bovine serum albumin (BSA) and to interfere with the induction of tolerance to human gamma-globulin (HGG). These adjuvant activities were shown to occur under conditions in which LPS could also function as a B cell mitogen. This positive correlation was established by utilizing two experimental situations in which LPS was non-mitogenic for spleen cells. Thus, on the one hand, it was found that LPS did not function as an adjuvant in C3H/HeJ mice, a unique strain whose spleen cells were also unresponsive to LPS-induced mitogenesis. On the other hand, in strains which did respond to LPS mitogenically, LPS failed to function as an adjuvant when it was chemically altered to reduce its in vitro mitogenic activity. A correlation was also observed between mitogenesis and the capacity of LPS to function as a specific immunogen i mice. In contrast to the sustained and prolonged plaque-forming cell response that was observed in mice whose spleen cells were also responsive to LPS-induced mitogenesis, the response was relatively transient in the C3H/HeJ strain. These results are discussed in view of the possible in vivo modes of action of LPS.


Rat cell surface antigens. I. Isolation and partial characterization of an Ag-B antigen. Rat major histocompatibility or Ag-B antigen was isolated from soluble extracts of lymphoid cells by affinity chromatography. The purified antigen was shown in SDS gel electrophoresis to have a molecular weight of 30,000 to 35,000 daltons. In addition, amino acid composition was found to resemble, generally, that of HL-A and H-2 antigens. It thus appears that the demonstrated homology between H-2 and HL-A extends also to Ag-B.


The antigenicity of myelin encephalitogenic protein: production of antibodies to encephalitogenic protein with deoxyribonucleic acid--encephalitogenic protein complexes. Rabbits immunized with monkey encephalitogenic protein (EP) complexed to deoxyribonucleic acid (DNA) formed complement-fixing antibodies that reacted well with EP from monkey, bovine, and human brain and the large EP of rat brain. Reactivity with the small EP of rat brain was much less. The antibodies, analyzed by the quantitative micro-complement fixation test, were of the 7S class, were produced in high titer, and failed to react with histone, lysozyme, or acidic liver extracts. Immunohistochemical studies revealed that the antibodies bound to central nervous system white matter from human, rat, guinea pig, and mouse as well as human peripheral nerve myelin. These findings demonstrate that the immunogenicity of EP is greatly enhanced by attachment to DNA and perhaps other negatively charged molecules. This method makes available anti-EP with which to investigate immunochemically the antigenic and conformational features of EP.


Suppression of reaginic antibody formation. I. Induction of hapten-specific tolerance. Reaginic antibodies to DNP and ovalbumin were indcued readily in B6D2F1 mice by a single intraperitoneal injection of 1 mug of DNP-ovalbumin suspended with 1 mg aluminum hydroxide in 0.5 ml of saline. The formation of anti-DNP reaginic antibody was completely suppressed by treatment of mice with a conjugate consisting of the hapten coupled to an isolgous nonimmunogenic carrier, viz., murine phi-globulins. However, this treatment did not affect the level of antibody formation to the carrier of the immunizing antigen. The induction of unresponsiveness was dose dependent, complete suppression being achieved with 1 mg of the tolerogen. The state of unresponsiveness could be maintained for prolonged periods (these observations were made over a period of at least 8 months) by repeated injections of the tolerogen at intervals of 2 months. More importantly, the state of unresponsiveness could be induced readily not only in normal, but also in sensitized mice, i.e., this treatment was capable of abrogating an ongoing reaginic response, and the suppression was immunologically specific. Hence this system appears to have a great potential for adaptation to the treatment of allergic diseases in man.


Suppression of reaginic antibody formation. II. The use of adoptive transfer system for the study of immunological unresponsiveness. By the use of the adoptive transfer system in syngeneic mice it was demonstrated that: i) spleen cells of animals tolerized by the i.v. injection of DNP8-MgammaG 1 month before adoptive transfer into x-irradiated recipients remained unresponsive when further challenged with the sensitizing antigen (DNP2.3-OA administered i.p. with Al (OH)3); by contrast, a significant response (typical of a secondary response) was obtained in recipients of cells from mice which had received only the sensitized antigen; ii) the anti-hapten IgE secondary response of primed cells was not affected by the presence of cells of tolerized mice when these two types of cells were administered and challenged together in x-irradiated recipients; iii) the transfer of spleen cells or serum of mice, tolerized 20 days before harvesting, into intact syngeneic recipients did not interfere with the normal development of the anti-DNP IgE response of the latter; iv) the anti-hapten responses of tolerized animals receiving spleen cells from normal or primed mice together with the sensitizing antigen resulted, respectively, in a primary or secondary response; (v) the unresponsive state of tolerized cells, achievkd by immunosuppression of the original donors with respect to the haptenic determinant, was maintained even after two serial exposures of the cells to the immunizing antigen in two consecutive adoptive cell transfers into x-irradiated recipients. These findings provide a strong basis for the interpretation that the hapten-specific tolerace induced in mice by treatment with DNP8-MphiG involved the elimination or inactivation of hapten-specific IgE-producing cells or the blockade of the receptors of these cells.


Two distinct chemotactic factor inactivators in human serum. The chemotactic factor inactivator (CFI) has been isolated from whole human serum by a combination of techniques including salt precipitation, anionic exchange, and gel filtration chromatography. Two inactivators have been obtained, a beta-globulin with a sedimentation velocity of approximately 7S and an alpha-globulin with a sedimentation velocity of approximately 4S. The former has a specificity for inactivation of the chemotactic activity associated with the C3 fragments, whereas the C5 chemotactic fragment is specifically inactivated by the alpha-globulin CFI. CFI in crude fractions of human serum is heat labile, time and temperature dependent for its activity, and pH dependent, expressing optimal activity at a pH range of 7.2 to 7.4.


Induction of tolerance to nucleic acid determinants by administration of a complex of nucleoside D-glutamic acid and D-lysine (D-GL). The administration of nucleosides coupled covalently to the copolymer of D-glutamic acid and D-lysine (D-GL) or to its stereoisomer, L-GL, induces a state of nucleoside (NUC)-specific tolerance in inbred SJL and BALB/c mice, irrespective of their immune status at the time of treatment. Such tolerance is characterized by the inability of treated animals to mount either primary or secondary, intact or adoptive, anti-NUC antibody responses following immunization with a highly immunogenic conjugate of NUC-Keyhole limpet hemocyanin. These observations have potential therapeutic importance in autoimmune processes involving anti-DNA antibody production.


Maturation of B lymphocytes in the rat. II. Subpopulations of virgin B lymphocytes in the spleen and thoracic duct lymph. Thoracic duct and spleen cells of normal (unimmunized) adult rats were fractionated according to size by 1 times G velocity sedimentation. Fractions were tested for their ability to restore the adoptive antibody response of irradiated hosts to horse spleen ferritin. A constant source of T cells (small numbers of unfractionated thoracic duct cells) was added to each fraction in order to monitor the B cell activity of the latter. Although large and small cell fractions of the spleen showed restorative activity, only the small cell fractions of the thoracic duct lymph showed activity. The turnover rate of the spleen cell fractions was determined by treating donors with high specific-activity 3H-thymidine for 48 hr before splenectomy. Rapidly dividing cells are preferentially killed by this treatment. The results suggest that a considerable proportion of large, intermediate, and small virgin B cells turn over within 48 hr. The cell surface of the various spleen cell fractions was examined for the presence of immunoglobulin (Ig) and a receptor for complement. The percentage of Ig-bearing cells in the large cell fractions was similar to the percentage of cells bearing IgM and a receptor for complement. However, the majority of Ig-bearing cells in the small cell fractions did not show the latter two surface markers. Experiments with the fluorescence-activated cell sorter showed that the large functionally active B cells bore surface IgM. The experimental findings suggest that there are subpopulations of virgin B cells in the spleen of the adult rat which differ with respect to size, migration pattern, turnover rate, and cell surface characteristics. The relationship of these cells to one another is discussed in the framework of an antigen-independent model of B cell maturation in the rat.


Mechanism of action of concanavalin A on human basophils. Concanavalin A (Con A) activates human basophils to release histamine. This reaction has great similarities to that of antigen-or anti-IgE-induced release. It is characterized by a two stage reaction: the initial stage is the binding of Con A to the cell and the second stage is a secretory process of the cell. Cells desensitized by incubation with excess amounts of Con A or by prolonged incubation in the absence of cations cannot be activated to release with antigen or anti-IgE. When the native (tetravalent) Con A molecule is modified by succinic anhydride to become divalent it is much less active in releasing histamine. There is wide variation in the response of cells from different individuals to Con A. However,  a close correlation exists between the histamine release induced by Con A and anti-IgE. Therfore, in genral, cells from allergic donors release more histamine with Con A than those from non-allergic individuals. Evidence is presented to demonstrate that Con A acts by binding to carbohydrate groups on the IgE molecule located on the basophil membrane. On a molar basis, IgE is 4000-fold more active than alpha-methyl-D-mannoside in inhibiting the Con A-induced histamine release. Succinyl Con A blocks the ability of Con A and anti-IgE equally well and to a lesser extent the ability of antigen to activate cells to release histamine.


Identification of a colon-specific antigen (CSA) in normal and neoplastic tissues. An antigen has been isolated from a human signet-ring cell carcinoma serially growing in hamsters, GW-39, by saline, PCA, or phenol extraction, and has been found immunologically identical to a similarly extracted substance in normal human or hamster colon. No other hamster or human tissues or cells were found to contain this antigen, for which reason we have termed it colon-specific antigen, or CSA. CSA has been found to be distinct from the major blood group-specific antigens and from othercolon tumor-associated antigens, such as CEA, CCA-II, and CCA-III. It thus seems that a colon organ-specific antigen can be synthesized by this particular human tumor system. Hamsters immunized with CSA could reject cheek pouch grafts of GW-39 tumors, and tumor rejection by these animals correlated with their anti-CSA antibody titers. Preliminary characterization of CSA suggested that it is a glycoprotein on the cell surface having a molecular size of 30,000 to 50,000 daltons. It is proposed that CSA may play a role in the diagnosis of mucin-producing adenocarcinoma of the colon and in ulcerative colitis.


Complement-mediated alteration of antibody specificity in vivo. The simultaneous injection of heterologous anti-EL4 lymphoma serum and complement results in the rapid disappearance of such antibody from the periphery of non-tumor bearing mice. However, this phenomenon is only observed when a complement source capable of mediating the lysis of EL4 cells sensitized with such heterologous antibody is used. This complement mediated enhancement of anti-tumor antibody absorption was observed in vivo for three strains of mice. Omission of complement or the use of genetically deficient complement sources resulted in no effect on circulating antibody titer when compared to the titer of heterologous anti-tumor antibody observed in the periphery when injected alone. Exogenous complement did not enhance the clearance of heterologous anti-tetanus toxin serum, thereby suggesting that the increased absorption of anti-EL4 in vivo is not related simply to the enhanced clearance of foreign gamma-globulin. Confirmatory evidence of the role of complement in altering anti-tumor antibody specificity in vivo was obtained in a guinea pig tumor model as well. The data suggest that anti-tumor serum shown to be relatively specific for the tumor cell gains additional specificity in the presence of functional complement and consequently manifests avidity for cross-reactive determinants previously thought to be unrelated.


IgA and IgA diphtheria antitoxin responses from human tonsil lymphocytes. Human tonsil lymphocytes were stimulated with diphtheria toxoid and then cultured in a Marbrook culture system so that antibodies could be measured in the culture supernatant. Specific antibodies were measured with excess radiolabeled antigen and antisera specific for each immunoglobulin class. Good IgG and IgA diphtheria antitoxin responses have been obtained and responding culture supernatants were shown to neutralize toxin. The relationship between antitoxin response in vitro and immunization of donors with toxoid was investigated. It was found that at least two immunizations after the age of 6 months were necessary to prime the tonsils for an in vitro antibody response. The IgG and IgA in culture supernatants were demonstrated by immunodiffusion and were measured by radioimmunoassay. By sucrose density gradient ultracentrifugation, it was shown that 40% of the IgA produced in the cultures was greater than 7S. Evidence was obtained that neither the IgA nor the specific IgA antitoxin bears secretory piece. It appears that human lymphocytes from tonsils produce polymer IgA in vitro without secretory piece.


Complement-mediated release of histamine from human leukocytes. Activation of either the alternative or classical pathway of complement generated a factor which induced release of histamine from both non-allergic and allergic human basophils. This factor probably is derived from the complement system since 1) its formation was associated with loss of C3 activity in human serum, 2) chemotactic factor, probably also a complement product, was generated simultaneously, 3) heat inactivation blocked its formation, 4) anti-C3 and anti-C5 blocked formation of the factor, and finally 5) anti-C5 inhibited the activity of the factor once it had been formed. It appears that both complement-mediated and allergen-mediated release of histamine from basophils are secretory, non-cytolytic pathways since both were maximal at 37 degrees C, required the presence of divalent cations, and were inhibited by theophylline. One consistent difference between these two mechanisms was noted: complement-initiated release of histamine occurred more quickly.


Application of the paired label radioantibody technique to detection and subtyping of hepatitis B surface antigen. A sensitive radioimmunoassay has been developed for the detection and subtyping, in regard to w and r specificities, of hepatitis B surface antigens (HBsAg). The binding of monospecific rabbit antibodies directed to either w or r determinant, labeled with 131I and 125I, respectively, by HBsAg was determined in the same experiment, taking advantage of the fact that these specificities are in general, expressed mutually exclusively. HBsAg in the test sample were bound to solid phase guinea pig anti-HBsAg antibody tube before the test. The ratio of percentage uptake of 125I anti-r to that of 131I anti-w for 28 normal sera without antigen was 0.69 0.18. The ratio for seven sera containing HBsAg with w determinant was less than 0.1, and that for 34 with r determinant was more than 10; 100-fold difference was noted between values of w and r antigens.


The hapten-carrier effect in teleost fish. The sea robin (Prionotus evolans), a representative teleost fish, was found to undergo the hapten-carrier effect, suggestive of T-B cell cooperativity, in vivo with dinitrophenyl as the hapten and bovine gamma-globulin or bovine serum albumin as carriers. In contrast to mammalian systems, the secondary anti-hapten response in the sea robin was found to involve exclusively high molecular weight antibody.


The diisopropylfluorophosphate inhibitable step in antigen-induced histamine release from human leukocytes. DFP inhibits early events in antigen-induced histamine release from human leukocytes. If added to cells 5 min or more after antigen it is ineffective. If added with antigen it can be removed at 5 min but release will still be inhibited. In contrast, ethylenediaminetetraacetate (EDTA) and 2 deoxyglucose (2DG) still inhibit the reactions when added 5 min after antigen. During incubation of leukocytes for 90 to 120 min at 0 degrees C they react with specific antigen since they subsequently release significant quantities of histamine after washing and reincubation at 37 degrees C without addition of antigen. Such priming at 0 degrees C is at least equivalent to priming for 2 to 4 min at 37 degrees C. During antigen priming at 0 degrees C the cells are not activated beyond the step in the release sequence which is inhibited by diisopropylfluorophosphate (DFP). This is apparent from the undiminished inhibitory activity of DFP on these cells. Furthermore, cells primed with antigen at 0 degrees C in the presence of DFP release as much histamine after washing and incubation at 37 degrees D as control cells primed in the absence of DFP. Incubation of leukocytes with specific antigen at 37 degrees C for 3 min resulted in significant but not quite complete priming for subsequent histamine release in the absence of antigen. Most of these primed cells were not activated beyond the step inhibitable by DFP. However, some had completed the entire sequence including the release of histamine while others had not released their histamine but were not inhibited by DFP from subsequent release. After 5 min incubation with antigen at 37 degrees C almost all leukocytes had progressed beyond the stage which is inhibited by DFP. Incubation of leukocytes at 37 degrees C with DFP but without antigen for up to 15 min followed by washing did not impair subsequent antigen-induced histamine release by these cells. Thus, DFP was inhibitory under these conditions only after antigen activation of leukocytes.


Detection of cell-bound immunoglobulins by a radioisotopic micro-mixed hemadsorption reaction with technetium-99m-labeled erythrocytes. The mixed hemadsorption (MHA) reaction detects antibodies reactive with cell surface antigens by means of antiglobulin-coated indicator erythrocytes. We have developed a radioisotopic modification which employs sheep erythrocytes (SRBC) that have been prelabeled with technetium-99m (99mTc), a high specific acitivity metastable gamma-emitter of short half life. The 99mTc MHA reaction was performed on human and murine cells cultured in Micro-test II plated with six replicate wells per serum dilution. Antibody activity in species-specific xenoantisera and mono- and polyspecific alloantisera was detected in high titer. The sensitivity of 99mTc micro-mixed hemadsorption was 2 times that of the visual assessment of mixed hemadsorption, 100 to 200 times that of the 125-I-mixed antiglobulin reaction and 500 to 1000 times more sensitive than indirect immunofluorescence. The assay system was applied successfully to confirm the species of origin of a panel of previously karyotyped human and mouse cell lines. Our results indicate that the 99mTc micro-mixed hemadsorption method is a rapid, sensitive, quantitative test for the detection of cell surface antigens and membrane reactive antibodies.


Studies on the cytophilic properties of human beta2 microglobulin. Human beta2 microglobulin is cytophilic for mouse, rat, and to a lesser extent guinea pig lymphocytes and polymorphonuclear leukocytes but not for human, rabbit, chicken, frog, and turtle cells, nor for any mature erythrocytes tested. Mouse splenic lymphocytes bind 5- 7-fold more beta2 microblobulin than thymocytes although hydrocortisone-resistant thymocytes resemble spleen cells in this regard. The beta2 microglobulin binding by B and T cells in mouse spleen is similar. The structure responsible for beta2 microglobulin binding to the surface of cells is distinct from the Fc receptor specific for aggregated IgG.


Tolerance to histocompatibility determinants in tetraparental bone marrow chimeras. Tetraparental bone marrow chimeras were produced by injecting lethally X-irradiated F1 hybrids with relatively high numbers of T-cell-depleted bone marrow cells from both allogeneic parental strains. The mice survival in excellent health and showed a stable, approximately 50:50 (parent:parent), lymphoid cell chimerism lasting for at least 7 mo after irradiation; regeneration of host-type hemopoietic cells was very limited. Thymus, lymph node, and thoracic duct lymphocytes showed specific unresponsiveness to host mixed leukocyte reaction (MLR) determinants. Similarly specific tolerance to H-2 antigens of host type was demonstrated in spleen and lymph node. No suppressor cells could be demonstrated in either system and blocking serum factors could not be found. The results suggest specific deletion of functional T cells reactive to host-type MLR and cell-mediated lympholysis determinants.


Suppression of the immune response by alpha-fetoprotein on the primary and secondary antibody response. Mouse amniotic fluid was shown to contain a noncytotoxic inhibitor of primary gammaM and secondary gammaM, gammaG subclass splenic plaque forming cells in vitro to SRBC. The suppressive effect was not abolished by exhaustive dialysis or by absorption of mouse amniotic fluid (MAF) with SRBC. Polyacrylamide gel analysis showed that dialyzed MAF was composed of three major protein components, transferrin, albumin, and alpha-fetoprotein (AFP). The selective removal of each of these patients from MAF by affinity chromatography suggested that AFP was the immunosuppressive substance in MAF. This conclusion was verified by the demonstration that pure AFP suppressed in vitro antibody synthesis in microgram quantities whereas equivalent amounts of normal mouse serum, transferrin, or albumin did not. Dose-response studies showed that the effect of AFP in the isolated form was equivalent to the suppressive effect of comparable amounts of AFP in MAF. gammaA and gammaG plaque-forming cell (PFC) responses were suppressed by a significantly lower concentration of AFP than was the gammaM PFC response. The degree of suppression watration of AFP than was the gammaM PFC response. The degree of suppression was dependent on the time at which AFP was added to the cultures; MAF added to antigen-stimulated cultures up to 24 h after initiation of cultures was immunosuppressive whereas similar additions of MAF at 48 h after initiation or later did not suppress. The duration of exposure of spleen cells to MAF in cultures without antigen necessary to achieve suppression of a subsequent primary immune response was determine-d to be approximately 8 h. The results suggest that AFP may have an immunoregulatry function. This has potentially important implications in the maternal-fetal relationship, the immune capabilities of the fetus and newborn, and in certain malignant and nonmalignant diseases in which AFP is elevated.


Localization of prostaglandin on the plasmalemma of rabbit sperm. Prostaglandin and prostaglandin receptors were localized on unwashed ejaculate rabbit sperm utilizing ferritin conjugated antibody to prostaglandin. Reaction was obtained over the acrosomal portion of the plasma membrane, with a greater aggregation over the apex of the head. The remainder of the head plasmalemma and flagellum showed no deposits of of reaction product. The reaction did not occur if the sperm were washed prior to incubation with the labelled antisera. However enhanced binding of ferritin was demonstrated if washed sperm were pre-incubated in PGE1. No reaction was seen in similar studies with human sperm.


Ultrastructure of an extremely thermophilic acidophilic micro-organism. A thermoacidophilic micro-organism, isolated from volcanic hot springs near Naples, was cultivated in vitro, and examined by electron microscopy in sections and after negative staining. The cells were almost spherical, with a diameter of about 0.7 to 1.0 mum. Their morphology was very primitive: the protoplasm was composed only of ground cytoplasm, ribosomes, and randomly distributed DNA strands. They were surrounded by a plasma membrane and by an extracellular coat about 20 nm thick which displayed a regular hexagonal pattern. Cell replication occurred by binary fission with median constriction during which a bipolar localization of nuclear material was observable. The morphology is compared with that of other known micro-organisms living in similar habitats.


An attempt to identify the intestinal receptor for the K88 adhesin by means of a haemagglutination inhibition test using glycoproteins and fractions from sow colostrum. The K88 antigen of Escherichia coli specifically adheres to the piglet intestinal cell; a solution of this antigen agglutinates guinea-pig red cells at 4 degrees C. The latter reaction was used as a model of the former, using inhibition of haemagglutination as an index of specific combination with the K88 adhesin. Inhibition was found with mucous glycoproteins and chemical modification of their heterosaccharide residues by mild acid hydrolysis, periodate oxidation or the Smith degradation procedure suggested that the terminal beta-D-galactosyl structure in a heterosaccharide sidechain of a glycoprotein might combine specifically with the K88 adhesin and inhibit haemagglutination. One serum glycoprotein (fetuin), after exposure of its subterminal beta-D-galactosyl residue, also inhibited haemagglutination, but high inhibitory activity was exhibited by some submaxillary glycoproteins in which this structure was absent or not prominent. It was concluded that in some cases inhibition of haemagglutination by glycoprotein was non-specific. No inhibition was found using glycosaminoglycans, glycogen or any simple sugar or glycoside. Sow colostrum was inhibitory but this was associated mainly with its gamma-globulin fraction. Some inhibitory activity was traced to a colostral glycopeptide fraction of low molecular weight but the smaller colostral oligosaccharides were not inhibitory; the composition of these components in sow colostrum is reported.


Experimental allergic optic neuritis (EAON) in the rabbit. A new model to study primary demyelinating diseases. In New Zealand albino rabbits the axons of the nerve fibre layer are myelinated over a long distance within the retina. Primary demyelination of these fibres was induced by sensitization of the animals to bovine cerebral white matter or myelin basic protein in complete Freund's adjuvant and challenge thereafter by intravitreous injection of basic protein (BP) or purified tuberculin (PPD). Production of segmental demyelination as a result of sensitization to brain-specific antigen (BP) and indifferent (PPD) antigen, shows that morphologically the same type of lesions can be induced by any antigen which will elicit cell-mediated immune reaction.


Brain tumor-scanning agents compared in an animal model. Sixteen radiopharmaceuticals for brain tumor localization have been compared in a mouse brain tumor model. A rating system is presented for such intercomparison. The rusults indicate that 111-in-chloride injected at pH 1.5 has the most favorable biologic characterisTICS FOR BRAIN TUMOR IMAGING.


A simple technique for making Diazochrome slides as teaching aids. Color pictorial slides are important for the learning process. They present a method which allows easy recognition of the subject without any effort to understand and appreciate it. However, the main purpose of this article is to describe a technique for the production of Diazochrome film for titles, charts, graphs, and diagrams. These slides are easy to produce, inexpensive, and time saving.


Hepatocarcinogenesis by diethylnitrosamine in rats fed high dietary levels of lipotropes. The effects of high dietary levels of lipotropes on the carcinogenic activity of diethylnitrosamine (DENA) in rats were studied. All animals given DENA, with or without a dietary supplement, developed hepatocellular carcinomas. The mean survival times of all groups of rats dying with hepatocellular carcinomas after DENA treatment were determined. Choline, betaine, and folic acid consistently exerted no significant effect on those mean survival times. In rats receiving approximately equal to 2 mg DENA/day, methionine administration led to a slight but significant increase in the mean survival time of the carcinogen-treated rats, whereas vitamin B12 significantly lowered the survival time; neither substance altered the mean survival times of those treated with only 1.0 mg DENA/day. On the other hand, ethanolamine decreased the mean survival times of rats given 1.0 mg DENA daily, but had no effect on animals receiving 2 ml/day of carcinogen. Dimethylthetin (sulfur analogue of betaine), methotrexate, lecithin, and cephalin exerted no effect on the carcinogenic activity of DENA. The administration of dimethylthetin and betaine along with DENA led to markedly increased liver weights in animals dying of hepatocellular carcinomas, when compared to liver weights of animals treated with DENA alone.


Regression of feline sarcoma virus-induced sarcomas in dogs. II. Immunologic investigations. The serial development of cell-mediated immunity (CMI), cytotoxic antibody activity, serum blocking activity, and virus-neutralizing antibody levels were monitored in vitro for beagle and mongrel puppies inoculated with feline sarcoma virus (FeSV) and were compared to in vivo histologic markers of regression of induced sarcomas. CMI developed rapidly and maintained a high level of in vitro activity throughout the tumor life-span. Cytotoxic antibody levels similarly rose rapidly to peak just before clinically detectable regression and then declined during most of the regression sequence. This suggested antibody fixation at the tumor site, correlating with the histologic finding of focal necrosis and neutrophilic infiltrates. Inactivation of antibody by circulating antigen with subsequent immune complex formation was a possibility. Levels of virus-neutralizing antibody in sera paralleled those of cytotoxic antibody; their relationship in the circulation was not clear, but each related to virus-determined antigenic specificities. Serum blocking activity rose rapidly, leveled off during most of the tumor life-span, and rose slightly during the last stages of regression. This partly explained the lack of in vivo tumor lymphoid infiltrates to correlate with the striking in vitro CMI. Blocking activity was also present, however, when lymphoid infiltrates were seen histologically. Thus in vitro-in vivo correlation was best for cytotoxic antibody, which suggested that antigen-antibody reactions involving neutrophil-mediated regression sequences were important in effecting tumor-cell destruction.


Argyrophilic carcinoids in two Syrian hamsters (Mesocricetus auratus). Argyrophilic carcinoids were found in 2 Syrian hamsters (Mesocricetus auratus). A solitary neoplasm was located in the glandular stomach of a 25.5-month-old animal that had ingested for 12 months laboratory chow to which arecoline and calcium hydroxide [Ca(OH)2] had been added. In the other hamster, a primary carcinoid of the pancreas had metastasized to the mesenteric lymph nodes and liver; this was in a 22-month-old animal that had ingested for 16 months chow containing added snuff and Ca(OH)I. It is not known whether the carcinoids were spontaneous or induced by the treatment. Argyrophilic carcinoids have previously been reported in only one other rodent species.


Antigenic characterization of type C RNA virus isolates of gibbon apes. Type C RNA viruses initially isolated from a lymphosarcoma of a gibbon ape and from a fibrosarcoma of a woolly monkey are very closely related immunologically. However, recent studies have shown that these viruses are distinguishable in a radioimmunoassay for the 12,000-molecular-weight polypeptide (p12) of the woolly monkey virus. In the present report, an immunoassay has been developed for the p12 polypeptide of the gibbon ape type C virus. This assay is shown to further distinguish the woolly monkey and gibbon ape viruses. In type-specific assays for the p12 polypeptides of these viruses, two new type C viruses isolated from gibbons in a second colony, characterized by high incidence of hemopoietic neoplasia, are immunologically distinguishable from the original gibbon ape virus. The p12 type-specific immunoassays described in the present report may be of importance in studying the natural history of these viruses and their relationship to tumors of primates.


Studies on reverse transcriptase of RNA tumor viruses. I. Localization of thermolabile DNA polymerase and RNase H activities on one polypeptide. Purified reverse transcriptase from avian myeloblastosis virus or Rous sarcoma virus consists of two subunits of average mol wt of 100,000 and 60,000. The lower-molecular-weight subunit, alpha, has been isolated from avian myeloblastosis virus, Rous sarcoma virus and a temperature-sensitive mutant of Rous sarcoma virus, LA337. Subunit alpha manifests both the DNA polymerase and RNase H activities associated with purified reverse transcriptase of avian RNA tumor viruses. The thermal inactivation of these enzymatic activities of alpha subunit from the wild-type virus. The results show that both DNA polymerase and RNase H activities associated with the alpha subunit of LA337 are five to seven times more thermolabile then the corresponding alpha subunit from the wild-type virus. It is concluded that (i) both the polymerase and nuclease activities reside on the same polypeptide chain, and (ii) at least the lower-molecular-weight subunit alpha is coded for by the viral RNA.


Effect of cytosine arabinoside on viral-specific protein synthesis in cells infected with herpes simplex virus. The relationship between viral DNA and protein synthesis during herpes simplex virus type 1 (HSV-1) replication in HeLa cells was examined. Treatment of infected cells with cytosine arabinoside (ara-C), which inhibited the synthesis of HSV-1 DNA beyond the level of detection, markedly affected the types and amounts of viral proteins made in the infected cell. Although early HSV-1 proteins were synthesized normally, there was a rapid decline in total viral protein synthesis beginning 3 to 4 h after infection. This is the time that viral DNA synthesis would normally have been initiated. ara-C also prevented the normal shift from early to late viral protein synthesis. Finally, it was shown that the effect of ara-C on late protein synthesis was dependent upon the time after infection that the drug was added. These results suggest that inhibition of progeny viral DNA synthesis by ara-C prevents the "turning on" of late HSV-1 protein synthesis but allows early translation to be "switched off."


Lifetimes of mRNA molecules directing the synthesis of viral proteins in herpes simplex virus-infected cells. Possible variations in the functional lifetimes of herpes simplex virus type 1 mRNA molecules in infected HeLa cells were studied. As shown by the rate of decrease of radioactive amino acid incorporation into viral proteins after the addition of actinomycin, the average lifetime of early viral mRNA's are shorter than those for the late messenger species. In addition, when the viral proteins made after the addition of actinomycin were further analyzed by gel electrophoresis, it was found that messengers for individual viral proteins translated within the early or late time period also had some differences in their functional lifetimes. These results indicate that the synthesis of herpes simplex virus type 1 proteins during the replicative cycle is regulated in part by mechanisms controlling the functional lifetimes of viral mRNA's.


Characterization of DNA polymerase and RNA associated with A-type particles from murine myeloma cells. The RNA-dependent DNA polymerase present in intracisternal A-type particles from mouse myeloma tumor cells has been studied. This polymerase can use either endogenous A particle RNA or an exogenous synthetic polynucleotide [poly (rA)] as a template. The DNA reaction product is small (4S-10S) and over 90% of it hybridizes to A particle RNA, whereas up to 50% of it hybridizes to murine sarcoma-leukemia virus RNAs. The RNA isolated from purified A particles is generally of low molecular weight (5S-15S) but contains small amount of 70S and 35S components. These results suggest that A-type particles may be related to C-type oncornaviruses.


RNA of simian sarcoma-associated virus type 1 produced in human tumor cells. Simian sarcoma-associated virus type 1 propagated in human rhabdomyosarcoma cells exhibited characteristics typical of oncornaviruses but seemed to have several aberrant properties. It had a buoyant density of 1.14 g/cm3, had RNA-dependent DNA polymerase activity, seemed to be labile to high salt concentrations, and contained little 50 to 60S RNA but relatively large amounts of human ribosomal RNA. In addition to 50 to 60S RNA, purified virions contained smaller RNA molecules with sedimentation coefficients of 28 to 30S, 18 TO 20S, and 4 to 10S. Unlike the 50 to 60S RNA species, the smaller virion-associated RNAs lacked polyadenylic acid, and the 28 to 30S RNA had an average base composition similar to that of human ribosomal RNA. Upon heat denaturation, the native 50 to 60S RNA genome yielded polyadenylic acid-containing 28 to 30S subunits that degraded in to 18 to 20S molecules upon further heat treatment. The 50 to 60S viral RNA had a guanine plus cytosine content of 56%.


RNA-dependent DNA polymerase activity of RNA tumor viruses. V. Rous sarcoma virus single-stranded RNA-DNA covalent hybrids in infected chicken embryo fibroblast cells. RNA-DNA covalent hybrids containing viral RNA have been isolated from nuclear fractions of Rous sarcoma virus-infected chicken embryo fibroblast cells shortly after virus infection. The formation of covalent hybrid structures depends upon a functional reverse transcriptase in vivo, since its appearance in cells is temperature dependent when infected with Rous sarcoma virus mutant LA335, which contains a temperature-sensitive reverse transcriptase.


Binding properties of avian myeloblastosis virus DNA polymerases to nucleic acid affinity columns. A new method for the analysis and purification of the RNA-directed DNA polymerase of RNA tumor viruses has been developed. This nucleic acid affinity chromatography system utilizes an immobilized oligo (dT) moiety annealed with poly (A). The alpha and alphabeta DNA polymerases of avain myeloblastosis virus bound effectively to poly (A) oligo (dT)-cellulose. Alpha DNA polymerase did not bind effectively to poly (A) oligo (dT)-cellulose, poly (A)-cellulose, or to cellulose. Alphabeta bound to oligo (dT)-cellulose and cellulose at the same extent (approximately 30%), indicating that this enzyme did not bind specifically to the oligo (DT) moiety only. However, alphabeta bound to poly (A)-cellulose two to three times better than to cellulose itself, showing that alphabeta could bind to poly (A) without a primer. Alphabeta DNA polymerase also bound to poly (C)-cellulose, whereas alpha did not. These data show that the alpha DNA polymerase is defective in binding to nucleic acids if the beta subunit is not present. Data is presented which demonstrates that the alphabeta DNA polymerase bound tighter to poly (A). oligo (DT)-cellulose and to calf thymus DNA-cellulose than the alpha DNA polymerase, suggesting that the beta subunit or, at least part of it is responsible for this tighter binding. In addition, alphabeta DNA polymerase is able to reversibly transcribe avian myeloblastosis virus 70S RNA approximately fivefold faster than alpha DNA polymerase in the presence of Mg2+ and equally efficient in the presence of Mn2+. alpha DNA polymerase transcribed 9S globin m RNA slightly better than alphabeta with either metal ion.


Bladder outlet obstruction caused by vaginal fibromyoma: the female prostate. Three patients with urinary difficulty were cured by surgical removal of a viaginal fibromyoma. The appearance of this benign tumor on an intravenous cystogram and cystoscopy as well as the clinical response to surgical removal bears similarity to the hypertrophied prostate in men. The best surgical technique for its removal is enucleation via a vaginal incision.


George Washington University technique for surgical correction of post-prostatectomy incontinence. We have described a procedure with long-term followup and modification for the correction of post-prostatectomy incontinence. The procedure involves the combined principle  of rigidly placed support under the urethra to which is attached an inflatable, adjustable pillow, allowing for fine control of the urethral resistance. Of the 22 patients treated 16 are completely dry, 4 have occasional dampness and 2 are failures. More significant is the fact that since the silicone gel pillow has been enclosed in the marlex strap we have had 6 successes., 1 partial successs and no failures. The complication rate is almost non-existent. No infections were encountered and no prostheses were removed for other than reoperation. We believe that the combination of bony fixation plus the adjustable pillow gives the best control of the troublsome problem of post-prostatectomy incontinence.


The significance of histological changes in serial prostatic biopsy specimens. Study of the histological appearance of randomly chosen areas of hyperplastic prostatic glands revealed that such areas are highly variable in appearance. Any such area of the size comparable to biopsy specimens does not give a representative picture of the entire gland. It is not possible from the appearance of single, serial biopsy specmens to draw any conclusions on the effect of hormonal treatment of the hyperplastic prostrate.


Morphogenesis of nodular hyperplasia--prostate. Step-section study was done on 161 prostates from medicolegal autopsies. Nodular hyperplasia occurred in 35-year-old subjects but increased in frequency with advancing age. Nodular hyperplasia originates as an early stromal nodule usually by the side of urethra. This nodule perhaps stimulates the duct in its close vicinity to proliferate and to bud into the solid nodule. Thereafter, stromal and epithelial elements proliferate to form either a glandular or a mixed nodule. A leiomyomatous nodule has been interpreted as a unilateral differentiation and maturation of stromal nodule to smooth muscle.


Polysaccharides of metaplastic mucosa and carcinoma of the gallbladder. The polysaccharide composition of the human gallbladder well was studied in carcinomas and metaplastic changes of various degrees, and the results obtained were compared with those for the normal material previously presented (Terho, T., and Laitio, M. Biochim. Biophys. Acta 338: 135, 1974). Elevated amounts of acid connective tissue polysaccharides (heparitin and dermatan sulfates as well as chondroitin 4- or 6-sulfate, or both, could be observed in carinomas. In histochemical stainings it was found that in carcinomas and in the two specimens classified as group III (containing the most extensive metaplastic changes at disposal), the intracellular mucin was mainly neutral or nonsulfated acidic. The amounts of sulfated mucin were relatively insignificant. This mucin polysaccharide material was isolated and its composition was determined. It was observed to be large polysaccharide material was isolated and its composition was determined. It was observed to be large molecular (approximate molecular sizes 1 to 2 times 10-6), and to be composed of fucose, galactose, glucosamine, and galactosamine as well as small amounts of sialic acid. The basic structure of these polysaccharides is thus similar to that of normal sulfated mucin. The almost total absence of acid groups, however, causes the polysaccharide material in question to stain in a manner identical with neutral mucin when investigated with histochemical methods. The carcinomas also contained some sulfomucin; its proportion, however, was small as compared with the amounts of nonsulfated acid and neutral mucin in biochemical characterization. A small molecular polysaccharide fraction, assumed to originate in membrane-bound glycoproteins, was isolated from the insoluble gallbladder tissue residue. The proportion of this fraction was larger in carcinomas than in normal material. This rise as well as the rise in the quantity of acid connective tissue polysaccharides is presumably due to the large number of cells in the carcinoma tissue as well as to fibrosis.


Incidence and nature of cytoplasmic hepatitis B antigen in hepatocytes. Hepatitis B antigen (HB Ag) in the hepatocytic cytoplasm is detected by immunofluorescence after reaction with fluoresceinated antiserum to HB Ag or by electron microscopy as numerous 20- to 30-nm. tubular and circular structures in dilated cisternae of excess endoplasmic reticulum. On light microscopy, these hepatocytes can be recognized because their cytoplasm has a ground-glass appearance and stains with Gomori's aldehyde fuchsin. Aldehyde fuchsin-positive ground-glass hepatocytes were detected in all 14 asymptomatic carriers of HB Ag and in 16 of 60 HB Ag-seropositive patients with chronic hepatitis, but not in HB Ag-seropositive acute viral hepatitis or in various other HB Ag-seronegative liver diseases. These cells are helpful in identifying on light microscopy HB Ag carriers and a portion of patients with HB Ag-positive chronic hepatitis. Nuclear HB Ag did not stain with aldehyde fuchsin. Nucleic acids were not detected in the ground-glass cytoplasm by special stains at the light or electron microscopic level. We suggest that the tubular and circular structures in the hepatocytic cytoplasm are coat material of the hepatitis B virus or virally coded host cell reaction product rather than the complete hepatitis B virus.


Identification of actin-like proteins in alcoholic hyaline by immunofluorescence. Immunofluorescence studies on human liver from alcoholic patients demonstrate that alcoholic hyaline (Mallory body) binds immune sera containing anti-actin antibodies. These findings, establishing that alcoholic hyaline is endowed with contractile proteins, strongly support the actin microfilament nature of the Mallory body.


Current status of the surgical treatment of truncus arteriosus. Persistent truncus asteriosus is now correctable surgically in patients with favorable anatomy. Given pulmonary arteries of reasonable size arising from any source, successful correction is possible so long as irreversible pulmonary vascular disease has not occurred. Although the majority of children with this defect demonstrate increased pulmonary blood flow, systemic-pulmonary artery shunts can be used. Also, banding of the pulmonary artery, followed subsequently by successful total correction, has been described. Recent reports of a few successful total corrections in infancy, performed with the aid of deep hypothermia and circulatory arrest, may modify the current approach. Although the majority of the reported corrections have involved aortic homograft reconstruction of the pulmonary artery, we strongly favor a synthetic prosthesis containing a heterograft valve. Based upon our clinical experience and this review of the literature, a suggested management protocol is presented.


Blood velocity at mitral valve during atrial arrhythmias in man. By means of the Doppler ultrasonic flowmeter catheter, phasic instantaneous left ventricular blood velocity was measured at the mitral valves of 184 conscious human subjects during atrial arrhythmias. Atrial extrasystoles and pacemaker-induced atrial tachycardias reduced peak mitral valve blood velocities in direct relation to shortened diastolic cycle length. Atrial fibrillation with irregular ventricular responses produced beat-to-beat alternations of peak mitral velocity, with smaller peak mitral blood velocities usually associated with shortened R-R intervals on the electrocardiogram. This report represents the first comprehensive description of instantaneous and continous phasic blood velocity at the mitral valve during atrial arrhythmias in man. It is concluded that atrial tachyarrhythmias exert an adverse influence on left ventricular filling velocities.


Initial effect of injury on ketone bodies and other blood metabolites. The arterial and venous concentrations of ketone bodies and other metabolites were measured in twelve adults, from 2 to 24 hours after injury. Two groups could be distinguished, with or without hyperketonaemia (defined as more than 0.2 mmol per litre) in the 2-hour blood-sample. In the hyperdetonaemic group the concentrations of alanine, pyruvate, and lactate and the urinary nitrogen excretion were lower throughout the first 24 hours than in the non-hyperdetonaemic group. These preliminary results indicate that hyperketonaemia after trauma is associated with decreased protein breakdown.


Neuropathy, M components, and amyloid.

Neuropathy, M components, and amyloid. Of eighty-seven patients with nonhereditary systemic amyloidosis, ten had evidence of peripheral neuropathy. There was median-nerve neuropathy due to infiltration of the flexor retinaculum with amyloid in six patients. Two patients had diffuse sensorimotor neuropathy and two patients had sensory loss without evidence of motor impairment--three of these cases had amyloid deposition demonstrated by nerve biopsy.  Neurological manifestations in all ten patients preceded other evidence of amyloidosis by 6 months-4 years (mean, 1 to 5 years). Neuropathy occurred in six of thirty-eight patients with primary amyloidosis and four of fourteen with amyloidosis associated with malignant B-cell dyscraias; and all patients with neuropathy had either a serum-M-component or Bence-Jones proteinuria. In contrast, neuropathy was not noted in any of the fifty-six patients in this series who had no evidence of a monoclonal gammopathy (thirty-five with secondary and twenty-one with primary amyloidosis).


The diving reflex used to treat paroxysmal atrial tachycardia. Induction of the diving reflex, by immersion of the face in cold water (2 degrees C) while the breath was held, converted paroxysmal atrial tachycardia to sinus rhythm within 15-35 seconds in seven patients (aged 22-66). Four had histories of heart attacks that had previously required vasopressor therapy, and two had been digitalised; three had no history of prior paroxysmal atrial tachycardia or heart-disease. The reported procedure, which is convenient, non-invasive, and can be self administered by the patient after brief instruction, may offer a useful adjunct to carotid-sinus massage and intravenous infusion of vasopressors for the treatment of paroxysmal atrial tachycardia.


The essential role of glucagon in the pathogenesis of diabetes mellitus. The following evidence suggests that diabetes mellitus may not be the simple consequence of relative or absolute insulin deficiency by itself, but may require the presence of glucagon: (1) relative or absolute hyperglucogonaemia has been identified in every form of endogenous hyperglycaemia, including total pancreatectomy in dogs; (2) insulin lack in the absence of glucagon does not cause endogenous hyperglycaemia, but when endogenous or exogenous glucagon is present, it quickly appears, irrespective of insulin levels at the time. These facts are compatible with a bihormonal-abnormality hypothesis, which holds that the major consequence of absolute or relative insulin lack is glucose underutilisation and that absolute or relative glucagon excess is the principal factor in the over-production of glucose in diabetes.


Plasma-high-density-lipoprotein concentration and development of ischaemic heart-disease. The body cholesterol pool increases with decreasing plasma-high-density-lipoprotein (H.D.L.) but is unrelated to the plasma concentrations of total cholesterol and other lipoproteins. This finding supports existing evidence that H.D.L. facilitates the uptake of cholesterol from peripheral tissues and its transport to the liver for catabolism and excretion. Plasma-H.D.L., is reduced in several conditions associated with an increased risk of future ischaemic heart-disease (I.H.D.), namely hypercholesterolaemia, hypertriglyceridaimia, male sex, obesity, and diabetes mellitus, while subjects with existing clinical I.H.D. have lower levels of H.D.L. than healthy subjects within the same community. It is proposed that a reduction of plasma-H.D.L. concentration may accelerate the development of atherosclerosis, and hence I.H.D., by impairing the clearance of cholesterol from the arterial wall.


Psychogeriatrics and community care.

Psychogeriatrics and community care. The meaning of community care, as at present organised for the elderly mentally ill, is examined critically. It is argued that often the phrase is used to conceal the deficiencies of traditional institutional facilities, for which community care is apt to be regarded as a cheap substitute. It is suggested that it would be beneficial to have available true community care, capable of providing a full-time service to patients in their own homes, in preference to institutional facilities. Though such a scheme would benefit patients, it would have implications for the community discordant with present official attitudes.


Patient's assessment of the result of surgery for peptic ulcer. The assessment of the result of surgery for peptic ulcer is based on doctor-determined criteria. Failure to distinguish one operation as being better than another may be because these criteria do not include the patient's rating of outcome. A questionnaire based on patients' descriptions of the quality of result has been tested in 63 patients. Preliminary figures show that success in the opinion of the patient does not necessarily mean absence of symptoms, while failure is due more to psychosocial than physical factors. A logical step in the planning of controlled trials is preoperative control of admission to the trial.


Mechanical deficiencies in scintillation counters, scanners, and gamma cameras. After an accident involving a scintillation counter, the machanical integrity of scintillation counters and their stands, rectilinear scanners, and gamma cameras was examined. An alarming proportion of these divices were found to be poorly designed with consequent danger to patients and staff. This class of equipment should be covered by standards incorporated in a comprehensive hospital technical memorandum.


Epidemic viral enteritis in a long-stay children's ward. Two outbreaks of acute gastroenteritis occurred in 1974 in a long-stay children's ward. Electron microscopy demonstrated rotaviruses in faeces from the affected children in the first outbreak, and adenoviruses in faeces from affected children and a nurse in the second outbreak. The illness in both outbreaks was very mild; but the diarrhoea associated with rotavirus infection usually lasted 5-8 days (in one patient it lasted for 28 days) and sometimes started with vomiting; whereas the adenovirus-associated diarrhoea lasted only 2-4 days and was not associated with vomiting. Neither the rotaviruses nor the adenoviruses could be established in tissue-culture.


New approach to management of intracranial aneurysms. In six cases an attempt was made to relieve the tension on intracranial aneurysms by temporarily clamping the internal carotid artery in the neck, so as to increase the expansibility of the artery. This approach was based on the concept (or "A principle") that haemorrhage is caused by the aneurysm having to bear the full force of systolic pulse pressure when atherosclerosis prevents this pressure being taken up by the normally expansile arterial wall. Follow-up has been fairly short, but the preliminary findings in four of the six patients are encouraging. More attention must be paid in the future to the significance of atherosclerosis in the onset of bleeding from intracranial aneurysms and the incidence of postoperative problems. The argument that atherosclerosis permits the transmission of the systolic pulse directly to the aneurysm wall requires further investigation. The earlier pathological signs of atherosclerosis must receive greater attention, and post-mortem study of the walls of arteries in immediate juxtaposition to aneurysms with high-powered magnification is required.


Effects of long-term practolol therapy on plasma-lipids after acute myocardial infarction. In a double-blind controlled trial lasting 12 months, long-term practolol therapy had no significant effect on plasma-lipid concentrations in twenty patients who had had acute myocardial infarction.


Salmonella typhimurium resistant to silver nitrate, chloramphenicol, and ampicillin. A strain of Salmonella typhimurium appeared sequentially in three patients in a burn unit, and epidemiological study suggested the occurrence of person-to-person spread. This organism was responsible for both colonisation and invasive infection in these patients whose burn surfaces were receiving topical treatment with 0.5% silver nitrate (AgNO3) solution. The antibiotic and metal ionsusceptibility pattern of this strain of S. typhimurium was unique and disturbing: resistant to silver nitrate, mercuric chloride, ampicillin, chloramphenicol, tetracycline, streptomycin, and sulphonamides. This pattern of multiple resistances could be transferred by invitro mating experiments to sensitive recipient strains of Escherichia coli and S. typhimurium. Further transfer of these resistances could be consumated between different strains of E. coli. A survey of other salmonella strains isolated from patients in this hospital without thermal burns did not reveal this pattern of resistance. Also, strains of S. typhimurium, isolated elsewhere and showing simultaneous resistance to both ampicillin and chloramphenicol, were not resistant to AgNO3 in vitro. The very real danger of this strain of S. typhimurium in burn units stems from its resistance to the two most effective antibiotics (ampicillin and chloramphenicol) available for systemic therapy; and this threat may be compounded through the selection effected by the widespread topical use of AgNO3 solutions and sulphonamide preparations on burned surfaces.


HL-A matching and kidney-graft survival. Scandiatransplant report. The follow-up of 523 kidney transplants with grafts from full-house donors shows a significant (p less than 0.02) correlation between the graft survival and the HL-A match grade. There was no indication in this study that kidneys from blood-group O donors do better in group O recipients than in non-O recipients. Neither was it possible to show any effect of matching for either of the haplotypes, HL-A1,8, HL-A3,7, or HL-A2,12.


Importance of a new virus in acute sporadic enteritis in children. In a year-long survey of children admitted with acute enteritis to hospital in Melbourne, Australia, an aetiological agent was found in approximately 75% of patients. During most months of the year a newly described virus belonging to the family Reoviridae was detected by electron microscopy of faecal extracts from more than 50% of patients. Recovery of this virus reached a peak (73%) during the winter. All age-group up to five years of age were susceptible to infection, which was occasionally fatal. Duration of infection was brief, with a probable incubation period of less than two days. Infections acquired in hospital were not uncommon. The name of this new virus is not decided. The name "duovirus" is proposed rather than "rotavirus" as previously suggested. There is already convincing evidence that this new virus will prove to be the most important aetiological agent of sporadic acute enteritis in young children.


Advanced diffuse histiocytic lymphoma, a potentially curable disease. Twenty-seven patients with advanced diffuse histiocytic lymphoma (reticulum-cell sarcoma) were treated with combination chemotherapy utilising nitrogen mustard (or cyclophosphamide), procarbazine, vincristine, and prednisone. Elven (41%) achieved a complete remission and only one of these has had a recurrence of tumour. The remaining ten complete responders were free of all evidence of tumour when last seen 26-105 months from the end of treatment. In contrast, all non-responders or partial responders have died. An interpretation of published survival data suggests that this virulent disease evolves quickly and is usally rapidly fatal if treatment is unsuccessful. Survival free of disease beyond 2 years from the end of treatment may be considered tantamount to cure. This definition of cure, previously applied only to patients treated with radiotherapy, seems applicable to patients who acheive complete remissions with modern drug treatment.


Heterozygous alpha 1-antitrypsin deficiency and cirrhosis in adults, a fortuitous association. Pi (protease inhibitor) genotype was determined in 394 healthy blood-donors, 132 adult patients with alcoholic cirrhosis, and 37 adult patients with cryptogenic cirrhosis. The frequency of the heterozygous genotype with a single allele Pi Z (heterozygous alpha 1-antitrypsin deficiency) was not different in blood-donors and in patients with cirrhosis. This finding suggests that the association of this heterozygous genotype with cirrhosis is not causal but fortuitous and that this heterozygous genotype does not increase susceptibility to cirrhosis due to other causes, in particular alcoholism.


Learned heart-rate control by a patient with a ventricular parasystolic rhythm. A 31-year-old woman with a ventricular parasystolic rhythm is described. The arrhythmia was always absent below a rate of 72 per minute, and always present over a rate of 106 per minute. Voluntary heart-rate control was learned using a biofeedback techinque, as a result of which she could increase her rate by 25 per minute and decrease it by 1-2 per minute. Both voluntary speeding and exercise brought on the arrhythmia, but later in the training she could reach a higher heart-rate before the arrhythmia appeared. Propranolol inhibited the arrhythmia only to the extent that is slowed the heart.


Minor calyces as primary pacemaker sites for ureteral activity in man. In isolated preparations of human upper-urinary-tract muscle in the organ-bath, calyceal and subcalyceal areas behave differently. Minor calyx preparations invariably exhibit immediate and extremely regular rhythmical contractions, in direct contrast to preparations of major calyx/pelvis which remain completely quiescent. Specimens of ureter do contract spontaneously but only after periods up to 1 hour. Smooth muscle of the minor calyces possesses excitatory alpha-adrenoceptors, as does that of the remainder of the upper urinary tract; but sensitivity of the minor calyx to alpha-adrenoceptor agonists is far greater than that of preparations from any other site. Furthermore, stimulation of intrinsic nerves can modify activity of the isolated minor calyx, whereas no such effect is observed in any area distal to the minor calyces. These physiological and pharmacological properties of the most proximal areas, considered together with the finding of structurally specialised smooth-muscle cells in this area, form the basis of a hypothesis at that minor calyces in multicalyceal kidneys act as a primary pacemaker sites.


Sudden death and sport.

Sudden death and sport. Of 21 sudden deaths in sportsmen, 18 were thought to be caused by heart attacks either during or after sport. There was firm evidence of ischaemic heart-disease in 9, strongly suggestive evidence in 7, but in 2 there was only suggestive clinical evidence. As a group, these subjects were characterised by (1) a mean age above thirty (above twenty-five for rugby players); (2) a family history of early heart-attacks; and (3) antecedent symptoms of chest pain or pressure in 9, fatigue or blackout in 4, and minor complaints in 2. Most were known to their medical practitioners. Psychological factors were thought to be important in 8. Doctors, players and referees should be aware that severe sporting exertion as in rugby football involves a risk which for most players is relatively minor, but in the minority predisposed to heart-attacks by family history, smoking, or age (as in referees) the risk is more serious. To reduce hazard of sudden death in exercise, players and referees should be warned against smoking and informed of the serious implications of the development of chest pain, pressure, or undue tiredness before, during, or after sport.


Gastric 'flu influenza B causing abdominal symptons in children. Influenza-B virus was identified in 102 children admitted to hospital during two epidemics in 1973 and 1974, enzbling the symptomatology of infection with this virus to be assessed in detail for the first time. Abdominal pain, often severe enough to require differentiation from acute appendicitis, emerged as a dominant symptom, especially in older children. Respiratory symptoms were often insignificant, although the lower respiratory tract was sometimes involved. Other symptoms in some children included convulsions and acute myalgia. The immunofluorescent method of virus diagnosis was found to be reliable for influenza B, except in a few cases ehere nasopharyngeal secretions were scanty, giving 97-5 percent copositivity with standard isolation techniques. The rapid result provided by immunofluorescence was helpful in clinical diagnosis and management and also in the control of hospital cross-infection.


Insulin responses to oral carbohydrate in true prediabetics and matched controls. The definition of prediabetes by genetic criteria alone has limitations since not all such subjects progress to overt diabetes.  Sequential oral glucose tolerance testing in a population has enabled the identification of 14 "true prediabetic" subjects with baseline two-hour plasma glucose levels smaller than 160 mg. per 100 ml. who subsequently developed unequivocal diabetes (two-hour plasma glucose level larger than 275 mg. per 100 ml.).  All but one were matched for baseline two-hour plasma glucose and relative weight with a subject whose glucose tolerance remained unchanged during a mean follow-up period of 4 years.  Fasting insulin levels and responses at 1/2, 1, and 2 hour sampling times were similar in both group and matched pair analysis at baseline.  No evidence was found that subjects destined to develop diabetes have either excessive or diminished insulin secretion.


Effect of continuous positive airway pressure on breathing pattern of infants with respiratory-distress syndrome. The effect of continuous positive airway pressure (C.P.A.P.) on the breathing pattern of ten newborn infants with respiratory-distress syndrome (R.D.S.) has been studied using an impedance pneumograph; arterial oxygenation improved and respiration, previously disorganized, became regular in both rate and depth.  Grunting usually ceased within 15 minutes of the start of C.P.A.P., and there was also on average a 30 percent increase in the respiratory-rate.  The rapidity with which the breathing pattern changed suggests a reflex mechanism.  Sudden reductions in airway pressure were frequently followed by apnoea, regular breathing restarting with the reintroduction of C.P.A.P.  These observations suggest that C.P.P.A. provides a respiratory drive in babies with R.D.S., possibly mediated through the Hering-Breuer inflation reflex.


Lymphocyte-dependent antibody cross-matching for transplant patients. Cadaver donor transplant patients who had a negative complement-dependent cytotoxicity cross-match test were tested in parallel with the specific donor at the time of transplantation by the lymphocyte-dependent antibody (L.D.A.) test. Transplants were performed on the basis of negative standard and long (2-5-hour) ment-dependent cross-match tests and the results of the L.D.A. test were withheld from the transplant centres. Of the 57 transplants tested, 49 had a negative L.D.A. result with 74 percent one-month function whereas 8 had a positive L.D.A. Result with 25 percent one-month function (P smaller than 0-02). When the results were further divided into methods of preservation of donor kidneys, among L.D.A.-Negative transplants, of the 20 kidneys preserved by Collins Sacks solution, 95 percent were functional at one month, whereas of the 28 kidneys preserved on the Belzer machine, 64 percent were functional at one month (P smaller than 0-05). The risk of the most hazardous period of transplantation (i. e., the first month) may be reduced drastically by simple preservation methods and the se of L.D.A. cross-matching.


Penetration of penicillin into human phagocytes containing gonococci. Human phagocytes that had ingested gonococci were incubated with increasing concentrations of penicillin in fresh human serum.  Viable counts on phagocyte deposits before and after incubation indicated penicillin penetration at high concentrations but not at lower concentrations which were sufficient to kill extracellular organisms.  With lower concentrations significant intracellular survival of gonococci occurred.


Functional role of cholesterol in infection and autoimmunity. Cholesterol binds to streptolysin O and related bacterial toxins.  In normal serum, only a fraction of the cholesterol attached to lipoprotein is available for binding, probably as a cholesterol-peptide complex formed during catabolic breakdown of the lipoprotein.  Cholesterol esterase produced by certain organisms--e.g., Staphylococcus pyogenes and Pseudomonas oeruginosa--augments this fraction both in vitro and in vivo.  Endogenous esterase similarly increases the amount of cholesterol-peptide complex, a mechanism which may be activated as a feedback process following binding  of toxin to the cholesterol component of the complex.  These complexes will thus supply a readily available means of binding bacterial toxins before antibody formation begins;  Cholesterol-peptide complexes, either alone or modified by binding to toxin, may function as autoantigens.  It is postulated that immune complexes so formed may be involved in atherosclerosis either by directly damaging vessels walls or by cross-reaction of antibody with cell-membrane-bound lipoproteins which equilibrate with plasma-lipoproteins.