Specificity of basophils and lymphocytes in cutaneous basophil hypersensitivity. Using a rosetting technique, it was found that the vast majority of basophils circulating in the blood or accumulating in the skin reactions of guinea pigs primed for cutaneous basophil hypersensitivity (CBH) lacked demonstrable specificity for sensitizing antigen, whether sheep erythrocytes, a soluble protein, or tumor cells. By contrast, one-third of cells teased from late skin reactions formed specific rosettes as did nearly 80% of circulating basophils in animals receiving repeated doses of whole sheep blood. Unreactive basophils teased from CBH reactions readily acquired rosetting capacity on exposure to immune serum. With regard to lymphocyte (and hence reaction) specificity, both CBH and classic delayed hypersensitivity (DH) reactions exhibited a high degree of carrier specificity when dinitrophenyl-conjugates were used. Thus, in the hapten-carrier combinations examined thus far, the antigen skin test requirements for both CBH and DH have been identical and are those required for inducing an active lymphocyte response. These findings indicate that control mechanisms other than homocytotropic antibodies must be sought to explain the accumulation and behavior of basophils in CBH reactions and, coupled with other data, suggest that lymphocytes and/or their products are likely candidates for this role.


Anti-receptor antibody. I. Isolation and characterization of the immunoglobulin receptor for phosphorylcholine. We have demonstrated a phosphorylcholine-binding protein in lysates from radioiodinated splenocytes of immunized mice. Immunoprecipitation with anti-receptor antibody of lysates from splenocytes obtained from animals undergoing a primary response or from mice immunized 4 to 6 months earlier with antigen demonstrated that this protein contained H and L chains. Therefore, we have isolated an antigen-specific receptor from the surface of spleen cells from both immunized and "memory" animals. This receptor Ig comprises approximately 10% of total cell surface Ig.


Anti-receptor antibody. II. Induction of long-term unresponsiveness in neonatal mice. We have examined the ability of anti-receptor antibody (ARA) to induce specific unresponsiveness to the hapten, phosphorylcholine (PC), in neonatal and adult mice. When ARA is given to adult mice, suppression is of short duration. Cells from such mice are responsive in vitro, indicating that suppression in vivo is probably due to blockade of receptors by persisting ARA. ARA given to neonatal mice induces long-term unresponsiveness. The mice apparently have decreased numbers of PC-responsive cells, since cells from such mice are unresponsive both in vitro and in adoptive transfer. Furthermore, cells from neonatally suppressed animals do not suppress the response of normal cells either in vitro or in adoptive transfer, indicating that unresponsiveness is most likely not due to active suppression. We therefore conclude that ARA given to neonates depletes the clone of receptor-bearing cells at the time ARA is given. Clonal depletion may result from antibody-dependent cell mediated cytotoxicity.


Control of histamine release: effects of various conditions on rate of release and rate of cell desensitization. Histamine release from rat mast cells by dextran and by antigen under various conditions was studied in terms of the rate of release and the duration of release, which together determined the total amount released. Reducing the concentration of the releasing agent decreased the rate of release without greatly affecting its duration. Reducing the temperature also decreased the rate, but prolonged the duration. Under all of the various conditions, release stopped as the cells became desensitized to the releasing agent. It appeared, therefore, that the rate of desensitization controlled the duration of release.


Genetic control of the immune response to the H-2 associated Slp alloantigen in the mouse. A survey of sixteen standard inbred and congenic resistant strains of mice reveals that the ability to mount an immune response to the Slp allotype is associated with the H-2 type of the recipient. Strains carrying the H-2f, H-2k, and H-2q haplotypes are able to produce specific antibody whereas strains of the H-2b haplotype are non-responders. Analysis of F1-hybrids and four informative intra-H-2-recombinants demonstrates that the ability to respond to the Slp allotype is controlled by a dominant gene associated with the K end of the H-2 complex.


Detection of membrane-associated antigens on lymphoid cells by antibody coupled to staphylococcal protein A. A simple technique is described for the detection of membrane-associated antigens on lymphoid cells. It is based on the observation that the protein A component of staphylococci binds to the Fc pieces of IgG molecules. Lymphocytes from various sources (mouse, rat, and human tissues) were incubated with hyperimmune antisera directed against surface determinants. Subsequent treatment with a suspension of staphylococci containing protein A permitted visualization of both the presence and distribution of determinants on the cell membrane. The method had comparable sensitivity to the fluorescent sandwich technique and could be used to detect a variety of membrane antigens on both T cells and B cells.


Complement-induced histamine release from human basophils. I. Generation of activity in human serum. The incubation of zymosan, endotoxin, or immune aggregates with normal human serum activates a factor which induces release of histamine from autologous basophils. The reaction can be divided into two steps: in the first, complement must be activated and in the second, the histamine-releasing factor interacts with basophils. The generation of histamine-releasing activity in serum occurs at 17 to 37 degrees C but not at 0 degrees C, is inhibited by heating the serum at 56 degrees C for 30 min, or by the addition of EDTA to the serum. Once generated, the histamine-liberating activity is stable to heating at 56 degrees C for 30 min. Gel filtration of the activated serum demonstrated that this factor eluted in the same region as a factor with chemotactic activity. Both factors have a molecular weight of about 16,000 daltons and their activities were inhibited by antibody to human C5. This is therefore a pathway for histamine release by C5a where the activation of the basophil is unrelated to the membrane bound IgE.


A radioactive hapten-binding assay for measuring antibodies to the pentapeptide determinant of peptidoglycan. A major portion of the humoral immune response to peptidoglycans is directed against the non-cross-linked pentapeptide side chains of these ubiquitous bacterial antigens. At present, no specific and sensitive assay for pentapeptide antibody determination is available. Therefore, a radioimmunoassay has been developed which employs the synthetic pentapeptide hapten L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala, labeled by the active ester method of Bolton and Hunter to high specific activities (6.74 to 18.18 muCi/mug) with 125I, and used as a reagent for measuring pentapeptide antibody. A-variant streptococcal antisera, known to contain pentapeptide antibodies as shown by quantitative precipitation, would bind more than 95% of the radiolabeled hapten in contrast to 2 to 3% by preimmune rabbit sera. Specificity of the binding reaction was demonstrated by inhibition experiments imploying various synthetic oligopeptides related or unrelated to the pentapeptide in the radioimmunoassay. Binding curves established with serial dilutions of peptidoglycan antiserum were linear from 15 to 500 mug/ml of antibody permitting pentapeptide antibody measurement within this range. Comparative data on pentapeptide antibody determinations by quantitative precipitation and radioimmunoassay are given and the time course of the production of this antibody in 14 rabbits hyperimmunized with A-variant streptococcal vaccine is reported.


In vitro studies on allotype suppression. II. Regulation of antibody synthesis by anti-allotype serum. The regulatory effects of rabbit antibodies specific for light chain determinants (b locus) on the formation of rabbit serum immunoglobulins have been studied in an in vitro system which measures of the response of unprimed rabbit spleen cells to solubilized T2 phage antigen. Treatment of spleen cells from b4b4 rabbits with anti-b4 serum, which was either incorporated into the culture medium or  employed in appropriate pulse treatment of the cells before culture, prevented the formation of T2 neutralizing antibodies by such cells. Spleen cells of heterozygous (b4b5) rabbits formed anti-T2 antibodies which could be shown to be divided  between the b4 and b5 specificities. Incorporation of anti-b4 or anti-b5 serum into the culture medium suppressed the specific anti-T2 response and, except in the instances noted in the text, did not significantly change the level of T2 neutralizing antibodies marked with the alternate allelic determinant. These findings are discussed in the light of the compensatory formation of an alternate immunoglobulin type which occurs during allotype suppression in vivo.


Studies on tolerance induction in vitro. I. Production of immunologically tolerant rabbit spleen cells and the transfer of tolerance to untreated cells. Tolerance was induced in rabbit spleen cells by incubation with solubilized T2 phage (S-T2)2 at 37degrees C. Spleen cells thus treated maintained normal responsiveness to an unrelated antigen, S-SP82. Transfer of tolerance was demonstrated in in vitro in that the addition of washed tolerant cells caused suppression of the response of untreated cells to an immunogenic dose of S-T2. Evidence is presented that this suppression is not due to the transfer of tolerogenic quantities of antigen. Spleen cell populations depleted of adherent cells were still capable of being made tolerant and of transferring tolerance.


Studies on the control of antibody synthesis. IX. Effect of boosting on antibody affinity. The effect of boosting on antibody affinity was studied in a haptenic system. Generally, boosting results in the prompt synthesis of high affinity anti-hapten antibody. However, repeated boosting frequently leads to a decrease in the amount and affinity of the serum antibody. Repeated boosting with hapten on a carrier different from that used for priming selectively stimulates synthesis of the highest affinity anti-hapten antibody and does not result in a decrease in affinity. Priming with soluble antigen without adjuvants results in the synthesis of low affinity antibody. After such priming, boosting stimulates low affinity antibody synthesis and repeated boosting leads to a moderate increase in antibody affinity.


Presence of thymic antigen on rabbit basophils. A goat heteroantiserum specific for a rabbit thymus lymphocyte antigen (RTLA) reacted with rabbit basophils resulting in the release of histamine. This activity was equally absorbed out by thymocytes or basophils. Absorption with either of these cell types also resulted in equal loss of thymocytotoxicity. No effect in either system occurred after absorption with either granulocytes or rabbit fibroblasts. These results could not be explained by the presence of immune complexes or aggregated globulin present in the RTLA antiserum. Further, the RTLA anti-serum had no anti-IgE activity, as demonstrated by its lack of reactivity with mast cells that were otherwise capable of releasing histamine normally after challenge with antigen. We have thus shown a basic difference between mast cells and basophils. We conclude that the rabbit basophil may bear a thymic marker.


Induction of blood eosinophilia by pulmonary embolization of antigen-coated particles: the relationship to cell-mediated immunity. A method of immunization has been described which consists of the i. v. injection into rats of large latex particles coated with human gamma-globulin (HGG). The coated particles embolize the pulmonary capillary bed and induce blood eosinophilia, which correlated strongly with the development of lymphocyte responsiveness to HGG in vitro. Sequential histologic studies of the lungs including ultrastructural analysis suggest a delayed hypersensitivity reaction to the coated particles. These studies support the concept that the induction of blood eosinophilia may be mediated by T lymphocytes.


Determination of antibody-hapten association kinetics: a simplified experimental approach. A simplified method has been developed for the determination of antibody-hapten association kinetics that permits the study of high affinity interactions with second order forward rate constants of the order of 10-7 to 10-8 M-1 sec-1. Use of tritiated haptens of high specific activity and antibodies of high affinity allows reactions to be run at initial hapten and antibody concentrations of the order of 10-9 to 10-10M, well below the level at which mixing becomes the rate-limiting step. Separation of antibody-bound from free hapten by the use of dextran-coated charcoal can be carried out with sufficient rapidity (2 sec) that the systems under investigation are not appreciably disturbed. With this technique, the association of 3-H-ouabain with rabbit ouabain-specific antibody was found to occur with a rate constant of 0.8 times 10-7 M-1 sec-1, similar to association rates  of dye haptens with antibodies of substantially lower affinity. The ratio of this association rate constant to the independently determined dissociation rate constant was 5.4 times 10-9 M-1, in satisfactory agreement with a ko value of 3.5 times 10-9 M-1 determined by Sips analysis of data obtained under equilibrium conditions. This approach should be applicable to the direct kinetic assessment of numerous high affinity antibody-hapten systems of current interest.


Role of the eosinophil in the allergic reactions. I. EDI-an eosinophil-derived inhibitor of histamine release. An inhibitor of histamine release was found to be associated with the human eosinophilic leukocyte. This eosinophil-derived inhibitor (EDI) was released from eosinophil-rich fractions upon sonication or interaction with immune reactants (specific allergens or anti-IgE). EDI was found to exert its inhibitory action at the target cell level by increasing the intracellular levels of cyclic-AMP. Preliminary electron microscopic studies show the presence of IgE on the eosinophilic leukocyte and it is suggested that the allergen or anti-IgE-induced release of EDI might be due to a direct interaction of these immune reactants with the eosinophil-bound IgE antibody. The results also suggest that by virtue of liberating a histamine release inhibitor such as EDI, the eopsinophil assumes a modulating role in the allergic inflammatory reaction.


Role of the eosinophil in the allergic reactions. II. Release of prostaglandins from human eosinophilic leukocytes. Human eosinophilic leukocytes respond to a variety of stimuli by liberating an inhibitor of histamine release. The active principle(s) in this eosinophil-derived inhibitor (EDI) was found to be a mixture  of acidic lipids of similar physiocochemical behavior and biologic activity to prostaglandins E1 and E2. Indomethacin, an inhibitor or prostaglandin synthesis, specifically blocked the immune release of EDI. The results indicate that stimulation of human eosinophilic leukocytes results in synthesis and release of prostaglandins E1 and E2 from these cells. These findings strongly support the hypothesis that the eosinophil assumes a modulatory role in the allergic inflammatory reaction. The ways by which, under specific antigenic (allergenic) stimulation, human eosinophils could modulate the events asssociated with the allergic response, through the released prostaglandins, are discussed.


Intracellular supravital stain delocalization as an assay for antibody-dependent complement-mediated cell damage. The ability of hamster SV40 tumor cells to concentrate Neutral Red into localized intracytoplasmic vacuoles and granules has been correlated with the ability of those cells to adhere and to replicate in tissue culture. Cells damaged by complement-fixing anti-tumor antibody and metabolic inhibitors first undergo delocalization of the dye, which appears as a diffuse stain throughout the nucleus and cytoplasm. More severely damaged cells lose the stain entirely, at a stage of progressive cell damage correlated with Trypan Blue uptake. A rapid and sensitive cytotoxic assay procedure has been based upon Neutral Red staining behavior, and the assay has been used to study antibody-dependent, complement-mediated cytotoxic anti-SV40 tumor activity in the sera of normal hamsters.


Radioelectrocomplexing: a counterelectrophoretic method for hapten--antibody interaction. We have described an application of a radioimmunoassay (RIA)method, known as radioelectrocomplexing (REC), which involves the anodal migration of antigen and the cathodal migration of antibody in agar electrophoresis. The agar is divided into zones of free antigen (DNP125I-HSA) and antigen bound with anti-DNP. Complete assays of anti-DNP can be performed in 2-4 hr since both immune complex formation and separation of free from bound antigen can be accomplished by electrophoresis in 60-90 min. Estimation of the weight of specifically-purified anti-DNP chicken antibodies in the nanogram range by REC is of the same order as the reported sensitivity of other RIA methods. The method was capable of demonstrating the higher avidity of the 17 S than 7 S antibody. Based on hapten inhibition the relative binding constants of DNP derivatives and anti-DNP were of the same order as reported from more definitive methods.


Immunofluorescent detection of hepatitis B antigen in paraffin-embedded liver tissue. Hepatitis B antigen (HBAg) has been demonstrated by the indirect immunofluorescent technique and by orcein staining in 20 liver biopsies fixed in Bouin's fixative and embedded in paraffin. The results were compared with those obtained previously by immunogluorescence on frozen sections of the same biopsies. Ten biopsies which were positive in frozen sections were also positive by immunofluorescence in parafin sections, whereas only six were positive by orcein staining. In orcein-stained sections, the cellular localization of HBAg was precisely in the same places as in the slides examined by immunogluorescence. The intessity of the fluorescence in paraffin sections was almost the same as in frozen sections. The localization of the antigen was histologically more precise in paraffin sections. Besides various advantages, indlucing aboidance of freezing aquipment and procedures, paraffin sections are more easy to handle and biopsies from distant hospitals can be processed. The advantages of the immunofluorescent test in comparison to orcein staining are its immunological specificity and higher sensitivity.


Hepatitis B surface antigen in blood donors: further observations. A survey of 128,000 volunteer blood donors from the Greater New York metropolitan area revealed that first-time male donors were positive for hepatitis B surface antigen (HBsAg) 2.5 times more frequently than were first-time female donors; Negroes and Mongols were positive four to 20 times more frequently than Caucasians. The ratio of ad to ay seemed to be higher in non-Caucasian antigen carriers than in Caucasian carriers. Among both Caucasians and non-Caucasians the rate of positivity declined after the age of 50. An excess prevalence of HBsAg was observed in donors with  the lowest level of education and in those with the highest level. HBsAg was detectable nine times less frequently among repeat donors than among first-time donors (0.2 vs.1.90 per 1,000). Detection of HBsAg was unrelated to ABO-Rh blood groups. Several mechanisms for these wide variations of antigen detection are possible.


Private specificities of H-2K and H-2D loci as possible selective targets for effector lymphocytes in cell-mediated immunity. Receptors of effector T lymphocytes of congeneic strains of mice do not recognize public H-2 specificities and react to private H-2 specificities only. This has been established with the use of three tests: direct cytotoxicity assay of immune lymphocytes upon target cells, specific absorption of the lymphocytes on the target cells, and rejection of skin grafts at an accelerated fashion. Immunization with two private H-2 specificities in the system C57BL/10ScSn leads to B10.D2 induces formation of two corresponding populations of effector lymphocytes in unequal proportion: a greater part of them is directed against the private specificity H-2.33 (Kb), while the smaller part is towards H-2.2 (Db) private specificity. These two populations of effector lymphocytes do not overlap, as demonstrated by experiments on their cross-absorption on B10.D2 (R107), B10.D2 (R101), B10.A(2R), and B10.A(5R) target cells, as well as on mixtures of R107 and R101 targets. Following removal of lymphocytes reacting with one of the private H-2 specificities, lymphocytes specific to the other specificity are fully maintained. A mixture of target cells, each bearing one of the two immunizing private specificities, absorbs 100% of the immune lymphocytes and is totally destroyed by them. It is suggested that H-2 antigens are natural complexes of hapten-carrier type, in which the role of hapten is played by public H-2 specifities and that of the carrier determinant by either private H-2 specificities or structures closely linked to them. Various models of steric arrangement of MHC determinants recognized by receptors of effector T lymphocytes are discussed.


Isolation from individual A/J mice of anti-rho-azophenylarsonate antibodies bearing a cross-reactive idiotype. Immuization of A/J mice with a KLH-p-azophenylarsonate conjugate induces the formation of antihapten antibodies, some of which share idiotypic specificity common to all recipients. The subpopulation carrying the idiotype generally comprises 20-70% of the total antibody content. Large quantities of antihapten antibody (occasionally over 100 mg) were obtained from individual mice through the induction of an ascites fluid. This facilitated isolation of antibodies with the cross-reactive idiotype by isoelectric focusing. Most of this subpopulation has pI values between 6.65 and 6.95 and essentially all is of the IgG1 subclass. Two peaks, near pI 6.7 and 6.9, were frequently observed. Upon refocusing, the protein artifact of the procedure, but indicates microheterogeneity. The antibodies in the two peaks were found to be idiotypically identical by measurements of cross-inhibition. Preliminary studies have indicated that it is feasible to initiate investigations of primary structure with antibodies from individual inbred mice.


Macrophage-lymphocyte interaction. II. Antigen-mediated physical interactions between immune guinea pig lymph node lymphocytes and syngeneic macrophages. The effect of specific antigen on the development of physical interactions between lymph node lymphocytes (LNL) obtained from animals which had been immunized to that antigen and macrophages was examined. We found that the presence of antigen, either limited to the macrophage () or free in the medium, profoundly increased the degree of ) or free in the medium, profoundly increased the degree of Mphi-LNL interaction observed. This enhanced interaction was dependent on the coincidence in the cultures of Mphi bearing antigen and LNL from animals specifically immunized to that antigen. Although antigen-independent interactions developed equally well between syngeneic and allogeneic combinations of lymphocytes and macrophages, antigen mediated interactions required that macrophages and lymphocytes be syngeneic. Prolongation of antigen-mediated Mphi-LNL interactions resulted in the induction of LNL DNA synthesis, initially involving those lymphocytes physically associated with antigen-bearing Mphi. These studies are interpreted to indicate that physical interaction between immune lymphocytes and antigen-bearing Mphi represents a morphological correlate of the functional activation of immune lymphocytes. Further, it is suggested that the physical events involved in lymphocyte proliferation may proceed sequentially from antigen-independent reversible binding of lymphocytes by macrophages to prolonged antigen-stabilized interaction eventuating in the triggering of specifically immune lymphocytes.


The oncornavirus glycoprotein gp69/71: a constituent of the surface of normal and malignant thymocytes. The oncornavirus related proteins associated with the surface of normal and malignant thymocytes were studied. Three virion-associated proteins (gp69/71, p45, p30) were associated with lymphoma cells from about 70% of the tumors studied. Two virion-associated proteins (gp69/71 and p45 were associated with normal thymocytes form some but not all strains of mice. In gp69/71- mice, conversion to the gp69/71+ phenotype accompanied leukemogenesis. An interesting difference in the apparent molecular size of virus related antigens of the 70,000 dalton size class was detected in lymphoma cells present in involved spleens as compared to involved thymuses. Mice infected as neonates with Scripps leukemia virus make antibody to gp69/71 and some make antibodies to molecules associated with the surface of their own tumors. The significance of the restricted presence of antigens coded for by the viral genome to the surface of some differentiated cells is discussed in reference to (a) the relationship between virion, leukemia associated, and differentiation dependent markers, and (b) the possible consequence to the host of having similar antigenic determinants on three independent structures with replicative potential (virus, normal thymocytes, and tumor cells).


Relation of GIX antigen of thymocytes to envelope glycoprotein of murine leukemia virus. Expression of Gix surface antigen on thymocytes is an inherited mendelian train of certain strains of mice. We report here the following new findings: (a) Gix antigen was found free in the serum of Gix+ mouse strains. (b) Expression vs. nonexpression of Gix antigen was invariably correlated with presence or absence of the group-specific antigen of Murine leukemia virus (MuLV) gp69/71 in the serum of mice of inbred and segregating populations. (c) Gix antigen could be removed from normal Gix+ mouse serum by precipitation with antiserum to MuLV gp 69/71. (d) Anti-gp69/71 serum was weakly cytotoxic for Gix+ thymocytes, and partially blocked the cytotoxic activity of Gix antibody for Gix+ thymocytes. (e) Purified AKR virus absorbed Gix activity, and disruption of the virions did not increase their absorbing capacity. These serological data indicate that Gix antigen is a constituent of gp69/71, the glycoprotein which is the major component of the MuLV envelope. On present evidence, Gix antigen is represented in intact virions and is probably accessible to Gix antibody.


Fetuin, an inhibitor of lymphocyte transformation. The interaction of fetuin with phytomitogens and a possible role for fetuin in fetal development. Fetuin, the bovine alpha-fetoprotein, contains glycopeptide sequences similar to those found on red cells. As a result, it is capable of strong physical interaction with the phytohemagglutinin isomitogens (H-PHAP) which possess two or more R (red cell binding) subunits as part of their tetrameric structures. Fetuin shows little or no interaction with L-PHAP, a phytohemagglutinin made up of four L subunits which also lack red cell affinity. Despite these differences fetuin is able to inhibit both H- and L-PHAP-induced lymphocyte transformation and is also capable of inhibiting the mitogenic effects of pokeweed mitogen, concanavalin A, antithymocyte antiserum, and the one-way mixed lymphocyte culture. In the case of L-PHAP, the inhibitory effect of fetuin is proportional to the intensity of the mitogenic stimulus. The inhibitory effects of fetuin upon lymphocyte transformation may result from perturbation or "blindfolding" of the cell membrane in a manner analogous to other immunosuppressive serum alpha-globulins. Alpha-Fetoproteins may play an immunoregulatory role during fetal development.


The specificity of cellular immune responses in guinea pigs. I. T cells specific for 2,4-dinitrophenyl-o-tyrosyl residues. Guinea pigs immunized with the hapten 2,4-dinitrophenyl (DNP) coupled directly to Mycobacterium tuberculosis of strain H37Ra (DNP-H37) show a variety of cell-mediated immune responses to DNP coupled to protein carriers. The cells responsible for this specific response are thought to be T lymphocytes for the following reasons: Guinea pigs immunized with DNP-H37 displayed delayed hypersensitivity reactions to several DNP-proteins and contact sensitivity to dinitrofluorobenzene. Peritoneal exudate lymphocytes (PELs) obtained from DNP-H37 immune animals respond to DNP-proteins with DNA systhesis and cause inhibition of macrophage migration. PELs are highly enriched in T lymphocytes and contain few immunoglobulin-bearing cells. Further depletion of immunoglobulin-bearing cells from this population does not diminish the in vitro proliferative response to antigen. Nitrophenyl conjugates of proteins lacking a paranitro group stimulated DNA synthesis poorly or not at all, indicating the importance of the paranitro group of DNP in antigen recognition by T cells in this system. In this respect, the specificity of T cells resembles that of DNP-specific antibody from the same animals. On the other hand, DNP conjugates of copolymers of glutamic acid and lysine and DNP conjugated to proteins via an interposed beta-alanyl-glycyl-glycyl spacer failed to stimulate DNA synthesis, although such compounds bind very efficiently to anti-DNP antibody. By contrast, DNP conjugates of synthetic polypeptide carriers containing as little as 7% tyrosine strongly stimulated DNA synthesis in DNP-H37 immune PELs. That the determinant responsible for this stimulation was DNP coupled to the hydroxyl group of tyrosine was shown by selective removal of DNP from tyrosine by thiolysis with 2-mercaptoethanol, which abolished their ability to stimulate T cells.


Heterogeneity of the BALB/c antiphosphorylcholine antibody response at the precursor cell level. Immune responsiveness to phosphorylcholine (PC) in BALB/c mice has been characterized by combining (a) usuage of highly sensitive radioimmunoassays for quantitation of antibody, heavy-chain class, and idiotype on a weight basis; (b) isolation of PC-specific B cells in fragment cultures; and (c) stimulation in a carrier-primed environment with the PC hapten coupled to carrier through a tripeptide spacer in order to maximize carrier recognition. The specificity and accuracy of the radioimmunoassays have veen verified by specific inhibition, lack of nonspecific binding, and excellent concordance of values for monoclonal antibody concentration obtained independently for Fab and idiotype content. The latter evidence also serves as strong confirmation of the monoclonality of in vitro monofocal responses as well as the preservation of the idiotype on antibodies of differing immunoglobulin classes. The results indicate that while B cells expressing the TEPC 15 idiotype predominate, other idiotypes may be represented by 2-50% of PC-specific precursors, and monoclonal antibodies even of the TEPC 15 idiotype are produced in both the IgM and IgG1 immunoglobulin classes. These findings are confirmed by the analysis of serum antibodies produced in carrier-primed mice immunized with hapten coupled through a tripeptide spacer, thus re-emphasizint the enhancement of primary responsiveness, particularly IgG1 production, by maximizing carrier recognition. The finding of idiotype diversity in the PC response, as well as diversity of expression in terms of quantity and immunoglobulin class of antibody synthesized by the clonal progeny of B cells within the TEPC 15 clonotype, emphasize the heterogeneity of the B-cell population both in terms of specificity repertoire and the physiological state of cells even within a single clonotype.


The immune response against myelin basic protein in two strains of rat with different genetic capacity to develop experimental allergic encephalomyelitis. After challenge with guiena pig basic protein (GPBP) Lewis (Le) rats, which are homozygous for the immune response experimental allergic encephalomyelitis (Ir-EAE) gene, developed positive delayed skin tests against GPBP and the 43 residue encephalitogenic fragment (EF); in addition, Le rat lymph node cells (LNC) were stimulated and produced migration inhibitory factor (MIF) when incubated in vitro with these antigens. In contrast Brown Norway (BN) rats, which lack the Ir-EAE gene, did not develop delayed skin tests to EF and their LNC were not stimulated and did not produce MIF when incubated in vitro with EF. These observations indicate that the Ir-EAE gene controls a T-cell response against the EF. Le rats produced measurable anti-BP antibody by radioimmunoassay after primary challenge. Although no antibody was detectable in BN rats by radioimmunoassay, radioimmunoelectrophoresis indicated that a small amount of antibody was formed after primary immunization. After boosting intraperitoneally, both strains of rat exhibited a rise in anti-BP antibody; which was greater in Le rats. In both strains of rat the anti-BP antibody reacted with a portion of the molecule other than the EF. Since EF primarily evokes a T cell response, it is suggested that the EF portion of the BP molecule may contain a helper determinant in antibody production.


Relationship between Fc receptors, antigen-binding sites on T and B cells, and H-2 complex-associated determinants. The relationship between H-2 complex-associated determinants, Fc receptors, and specific antigen-recognition sites on T and B cells was examined by binding and functional assays. The Fc receptor was detected by radiolabeled immune complexes or aggregated human IgG. Both these reagents selectively bound to B cells, not to T cells. When spleen cells, from mice primed to several antigens, were exposed to highly substituted radioactive aggregates, their capacity to transfer both a direct and indirect plaque-forming cell response to these antigens was abrogated. Addition of B cells, but not of T cells, restored responsiveness. Complexed Ig binding to Fc receptors was prevented by pretreatment of mixed lymphoid cell populations with antisera directed against membrane components on the same cell (e.g., H-2) and on other cells (e.g., theta). The lack of specificity of inhibition was thought to be due to the formation on cell surfaces of antigen-antibody complexes which would then attach to the Fc receptor during the incubation precedure. Specific blockade of the Fc receptor during the incubation procedure. Specific blockade of the Fc receptor however occurred when B cells were pretreated with the Fab fragments of anti-H-2 antibody. This was demonstrated autoradiographically and by inhibition of aggregate-induced suicide. The blocking activity of ante-H-2 Fab was removed by absorption with spleen cells from thymectomized irradiated mice but not with thymus cells of appropriate specificity. This suggested that the antibodies involved had specificity for determinants on the B-cell membrane distinct from those coded by the K or D end of the H-2 complex, and either absent from, or poorly represented on, thymus cells. Specific antigen-induced suicide of B cells was achieved simply by incubating the cells with radioactive antigen in the cold. T-cell suicide on the other hand required that the 125I-labeled antigen be presented to the T cells at 37 degrees-C on the surface of spleen cells from antigen-primed mice. Pretreatment of T cells with the Fab fragment of anti-H-2 antibody protected them from the suicide effect. By contrast no such protection of B cells could be achieved by this procedure. In other words H-2 (? Ir)-associated determinants may not only be in close proximity to the antigen-binding site on T cells but, in addition, may be involved in the effective operation of the receptor.


Structural, functional, and idiotypic characteristics of a phosphorylcholine-binding IgA myeloma protein of C57BL/ka allotype. An IgA phosphorylcholine (PC)-binding myeloma protein with IgCH allotypic determinants different from those of BALB/c mice is characterized. The myeloma, CBPC 2, was induced in the CB-20 strain of mice which is congenic to BALB/c but differs from it by carrying the A15 allotypic determinant of C57BL/ka mice. Sequence analysis of the CBPC 2 light chain through the first hypervariable region, as well as isoelectric point analysis, show that this chain is indistinguishable from that of T15, a PC-binding myeloma protein of BALB/c origin. The heavy chains of CBPC 2 and T15 differ by only two amino acids (positions 14 and 16) through the first hypervariable region. As measured by inhibition of precipitation, both CBPC 2 and T15 have the same specificity for PC, glycerophosphorylcholine, acetylcholine, and choline. In addition, CBPC 2 possesses the binding site-associated idiotypic determinant which is present on T15. However, like normal or induced C57BL/6 anti-PC antibody, it does not possess the nonbinding site idiotypic determinant.


Evidence for specific suppression in the maintenance of immunologic tolerance. Specific suppressor cells have been demonstrated in mice tolerant to the thymus-dependent antigen HGG. Transfer of normal thymocytes, normal spleen cells, or immune spleen cells into these tolerant mice did not restore immunocompetence to HGG. Furthermore, the transfer of tolerant spleen cells into normal recipients abrogated the response of these recipients to subsequent challenge with immunogenic HGG. Spleen cells removed from mice 5, 8, or 11 wk after the induction of tolerance specifically suppressed the response of normal spleen cells in an adoptive cell transfer system. The extent of suppression appears to be dependent upon how long after the induction of tolerance the cells were removed from the tolerant donors and how soon after transfer the recipients were challenged.


Antigen-specific T-cell factors in the genetic control of the immune response to poly(Tyr,Glu)-polyDLAla--polyLys. Evidence for T- and B-cell defects in SJL mice. The cellular basis of the genetic control of the immune response to poly(LTyr, LGlu)-polyDLAla--polyLLys [(T,G)-A--L] in SJL (H-2s, low responder) mice has been investigated using T-cell factors. Thymocytes of SJL origin were educated to (T,G)-A--L and tested for their ability to produce an antigen-specific factor capable of cooperating in vivo with bone marrow cells of either SJL or C3H.SW (high responder) origin. SJL T cells were found to be incapable of producing such a cooperative factor, in contrast with results previously obtained with C3H/HeJ (low responders) and C3H.SW strains. Moreover, SJL bone marrow cells did not produce an antibody response to (T,G)-A--L, even when combined with factor produced by high responder (C3H.SW) mice. Thus, both T and B cells appear to be defective in the SJL strain in the response to (T,G)-A--L.


Ultrastructure of thalamic neuronal inclusions in myotonic dystrophy. Recently, the presence of thalamic neuronal cytoplasmic inclusions in patients with myotonic dystrophy has been reported. At the ultrastructural level, the inclusions were described "containing a fibrillar material within a limiting membrane studded on its outer surface with ribosomes". We have studied the brain of a 48-year-old woman with myotonic dystrophy. Many neuronal inclusion bodies were found within the thalamus, and examined in the electron microscope. Inclusions were found to have an electron density resembling that of nuclear chromatin, were not membrane-bound, and had an internal structure composed of parallel alternating dark and pale lines. We confirm the previously-reported existence of cytoplasmic thalamic neuronal inclusions in myotonic dystrophy, but differ in our observations of their ultrastructural appearance, and note that these inclusions bear no resemblance to previously described inclusion bodies.


Neuritogenic and encephalitogenic properties of the peripheral nerve basic proteins. Two basic proteins, P1 of molecular weight 14,200 and P2 of molecular weight 12,300, purified from bovine peripheral nerve, were assayed for biological activity. The P1 protein is an exclusively neuritogenic agent, capable of producing clinical signs of experimental allergic neuritis (EAN) and histological abnormalities in the peripheral nervous system (PNS) of guinea pigs and rabbits, without any changes in their central nervous system (CNS). P2 protein, like the CNS basic encephalitogenic protein (BE), has combined neuritogenic and encephalitogenic activities, therefore it induces in these animals neurological signs and pathological evidence of EAN, as well as histological characteristics of experimental allergic encephalomyelitis (EAE).


The effects of the H1 and H2 antihistamines on "allergic" histamine release and its inhibition by histamine. Antigen-induced, IgE-mediated release of histamine from human basophiles is an in vitro model of allergic reacttions; it is blocked by extracellular histamine, presumably as a result of its ability to increase adenosine 3',5'-monophosphate (cyclic AMP) levels. The H1 antihistamines do not antagonize these effects of histamine but at approximately equal to 1 mM cause histamine release and at approximately equal to 0.1mM inhibit antigen-induced histamine release. The phenothiazine antihistamines are 10-30 fold more potent inhibitors than the rest; other tricyclic antidepressant drugs share this activity. The mechanism of this inhibition, which occurs in both the 1 degree and 2 degree stages of histamine release, is not known but it is not due to partial agonist activity since the anti-H1 drugs cause a significant fall in cyclic AMP levels. The anti-anaphylactic effects of the H1 antagonists probably play no therapeutic role but we suggest that drugs structurally similar to the phenothiazine antihistamines should be developed for clinical testing. The H2 antihistamines block histamine-induced inhibition of histamine release and the increase in cyclic AMP levels, but neither cause nor inhibit histamine release. The K-B values for the anti-H2 drugs (burimamide approximately equal to 5 muM); metiamide approximately equal to 0.5muM); are similar to those described for other H2 receptors.


Antibodies to fentanyl.

Antibodies to fentanyl. Immunization of rabbits with carboxyfentanyl-bovine gamma-globulin conjugate induced antibodies of high titers capable of binding with fentanyl.  The antibodies exhibited high average association constants (approximately 1 times 10-7 liter/mol) and were highly specific to fentanyl since no cross-reactivity was observed for a variety of test compounds including other opiates and analgesics.  Serological cross-reactivity was observed only for chemicals representing various portions of the fentanyl molecule.  These high-affinity, highly selective antibodies have been successfully employed in a radioimmunoassay for fentanyl.  Serum concentrations of fentanyl, after administration of 100 and 21 mug/kg/ i.v. in dogs, were determined by this radioimmunoassay.  The specificity of these antibodies as compared with the specificity of the pharmacological (opiate) receptor is also discussed.


Purification and characterization of the DNA polymerase and RNase H activities in Moloney murine sarcoma-leukemia virus. Two RNase H (RNA-DNA hybrid ribonucleotidohydrolase, EC 3.1.4.34) activities separable by Sephadex G-100 gel filtration were identified in lysates of Moloney murine sarcoma-leukemia virus (MSV). The larger enzyme, which we have called RNase H-I, represented about 10% of the RNase H activity in the virion. RNase H-I (i) copurified with RNA-directed DNA polymerase from the virus, (ii) had a sedimentation coefficient of 4.4S (corresponds to an apparent mol wt of 70,000), (iii) required Mn-2+ (2 mM optimum) for activity with a [3-h]poly(A)-poly(dT) substrate, (iv) eluted from phosphocellulose at 0.2 M KC1, and (v) degraded [3-H]poly(A)-poly(dT) and [3-H]poly(C)-poly(dG) at approximately equal rates. The smaller enzyme, designated RNase H-II, which represented the majority of the RNase H activity in the virus preparation, was shown to be different since it (i) had no detectable, associated DNA polymerase activity, (ii) had a sedmimentation coefficient of 2.6S (corresponds to an apparent mol wt of 30,000), (iii) preferred Mg-2+ (10 to 15 mM optimum) over Mn-2+ (5 to 10 mM optimum) 2.5-fold for the degradation of [3-H]poly(A)-poly(dT), and (iv) degraded [3-H]poly(A)-poly(dT) 6 and 60 times faster than [3-H]poly(C)-poly(dG) in the presence of Mn-2+ and Mg-2+, respectively. Moloney MSV DNA polymerase (RNase H-I), purified by Sephadex G-100 gel filtration followed by phosphocellulose, poly(A)-oligo(dT)-cellulose, and DEAE-cellulose chromatography, transcribed heteropolymeric regions of avian myeloblastosis virus 70S RNA at a rate comparable to avian myeloblastosis virus DNA polymerase purified by the same procedure.


Studies on reverse transcriptase of RNA tumor viruses III. Properties of purified Moloney murine leukemia virus DNA polymerase and associated RNase H. DNA polymerase was purified from a cloned isolate of Moloney murine leukemia virus (M-MuLV). Purified M-MuLV DNA polymerase, upon analysis by polyacrylamide gel electrophoresis, showed one major polypeptide of mol wt 80,000. Estimation of molecular weight from the sedimentation rate of the purifed enzyme in a glycerol gradient was consistent with a structure containing one polypeptide. M-MuLV DNA polymerase could transcribe ribopolymers, deoxyribopolymers, and heteropolymers as efficiently as did purified DNA polymerase from avian myeloblastosis virus (AMV). M-MuLV DNA polymerase, however, transcribed native 70S viral RNA less efficiently than did AMV DNA polymerase. Addition of oligo(dT) enhanced five to tenfold the transcription of 70S viral RNA by M-MuLV DNA polymerase. Purified enzyme also exhibited nuclease activity (RNase H) that selectively degraded the RNA moiety of the RNA-DNA hybrid. It did not degrade single-stranded RNA, single-stranded DNA, double-stranded RNA, and double-stranded DNA. M-MuLV DNA polymerase-associated RNase H acted as a random exonuclease. When [3-H]poly(A)-poly(dT) was used as a substrate, the size of the M-MuLV DNA polymerase-associated RHase H digested product was larger than the size of the digestion products by AMV DNA polymerase. The oligonucleotide digestion products could be further digested to 5'-AMP by snake venom phosphodiesterase, indicating that the products were terminated by 3'-OH groups. Alkaline hydrolysis of the oligonucleotide digestion products generated pAp, suggesting that M-MuLV DNA polymerase-associated RNase H cleaves at the 3' side of the 3',5'-phosphodiester bond. The ratios of the rates of DNA polymerase activity and RNase H activity were not significantly different in the murine and avian enzymes.


Renal protein metabolism in obstructive uropathy. Measurements of serum beta 2 microglobulin and creatinine concentrations were determined in 18 patients with lower urinary tract obstruction. Nine patients were azotemic on admission to the hospital. In 3 of these patients who had normal pre-treatment serum levels of beta 2 microglobulin, the correction of obstruction was followed by return of normal renal function. The remaining 6 patients with elevated serum beta 2 microglobulin concentrations on hospitalization, showed no significant improvement in renal function parameters in the post-obstructive period. These data suggest that determination of serum beta 2 microglobulin concentrations is helpful in predicting the return of renal function in azotemic patients with obstructive uropathy.


Programmed "trendscription." A new approach to electrocardiographic monitoring. A new approach to monitoring of episodic but frequently recurring arrhythmias has been developed, consisting of condensation of electrocardiographic information on a special drum recorder, a programmer that permits selection of intervals and durations for monitoring, and a telemetry unit for transmission of the electrocardiographic signal. We used this system to manage three selected cases.


[Certain practical problems of recognition and treatment of cardiac rhythm disorders]. Having briefly touched upon the problem of terminology and classification of arrhythmias the authors consider the diagnosis and clinical evaluation of some disorders, including the extrasystole (the significance of the extrasystolic interval), ectopic arrhythmias from the region of the atrio-ventricular junction (with simultaneous consistent or transent disruption of the intraventricular conduction), isolated atrial tachycardia, some variants of auricular fibrillation, flutter paroxysms, paroxysms of ventricular tachycardia, with continued auricular fibrillation in particular, and also the earlier described electrocardiographic phenomenon tentatively interpreted as sinistroatrial fibrillation with dextraatrial tachycardia, as well as major types of disrupted condution. The authors give a brief exposure of their views as to the principles of the treatment. Emphasis is placed on the importance of a comprehensive clinical approach to the diagnostic matters and to the evaluation of arrhythmias, as well as to the fundamental need to define more precisely the pathogenesis of the disturbed rhythm in a concrete patient so as to adopt an effective treatment.


[Prognosis of the effect of cardiac glycosides on the extrasystole in ischemic heart disease]. The results of a single intravenous injections of strophanthin or digoxin and of a course-wise medication with cardiac glycosides in acute and chronic ischemic heart disease are contrasted. Investigations were carried out in 64 patients under cardiomonitoring with an automatic extrasystoles count. In the absence of extrasystoles or infrequent ones the cardiac glycosides provoked frequent extrasystoles in 10 per cent of the patients. The antiarrhythmic effect was recorded in 1/3 of the patients with infrequent extrasystoles. In cases of frequent extra systoles the positive result of the glycoside test justifies anticipating a beneficial influence of digitalization on the heart rate. In all the cases the results of the glycoside test determine in a large measure the individual prognosis.


[Disorders of cardiac rhythm in chronic alcoholics]. The friquency and type of cardiac rhythm and conduction disturbances were studied in 200 patients with chronic alcoholism. Attention was payed to the character and duration of dipsomania. Patients with a long abuse period were shown to have auricular fibrillation in 10 per cent  of the cases. Patients with chronic alcoholism and particularly with rhythm disturbances demonstrated the decrease of potassium concentration in blood serum.


[Experience with the use of izoptina].

[Experience with the use of izoptina]. The effect of isoptin was studied in 271 patients with ischemic heart disease and various disturbances of the cardiac rhythm. Clinical, functional indices and coronarographic findings proved the drug to be highly effective. The mechanism of its action is discussed.


Palliative treatment of complete transposition of the great vessels: First clinical results of closed atrial septectomy with new septectome. Our initial method of closed atrial septectomy, used in the palliative therapy of transposition of the great arteries (TGA), failed to produce a technical success in 1 of 5 patients. Subsequently, we modified the technique by securing the septum on traction sutures and simplifying the introduction of the septectome. With the new method, we achieved technical success in all 13 patients so treated. In our total of 18 patients treated by closed atrial septectomy, 7 had TGA with no other cardiac anomaly, and none of the 7 died. Of the remaining 11 patients who had TGA and other cardiac anomalies and required procedures such as ductus ligation, pulmonary artery banding, and aorta-right pulmonary artery shunt in addition to septectomy, 5 died. In only 1 of them, death was caused by technical complications with our initial method. Of the remaining four deaths, two were due to pre-existing cerebral damage and two to complications of palliative procedures done in conjunction with atrial septectomy. Side-to-side anastomosis between the superior vena cava and right pulmonary artery, carried out successfully in 1 case, is suggested as an additional palliative measure to supplement atrial septectomy in TGA.


Alveolar lavage cytology in transplanted lungs. I. Staining methods and findings in dogs with autografts and allografts without immunosuppression. Alveolar lavages were performed repetitively on the normal and transplanted lungs of dogs that had recieved autografts or allografts without immunosuppression. One half of the lavage returns was fixed as a cytologic smear; the other half was subjected to semi-thin section or electron microscopic examination. Of the staining methods was used, the periodic acid-Schiff (PAS) and Giemsa techniques were best for differentiating and counting cells. The Ladewig technique was best for evaluating the presence and location of fibrin. After autotransplantation, the proportion of so-called alveolar marcophages increased, reached a peak in 4 to 7 days, and then returned to normal. Phagocytized fibrin increased for the first postoperative week, but not extracellular fibrin was ever observed. After allotransplantation, a progressive decrease in the proportion, size, and vacuolization of so-called alveolar macrophages was noted along with an increase in extracellular fibrin. Intracellular fibrin could be detected only up to the third day. These findings define adequate methods for preparing and staining material obtained from diagnostic alveolar lavages, and they suggest that the procedure may serve as an index of lung allograft rejection.


Congenital tricuspid stenosis treated by a palliative open operation. Report of a case. The case is presented of a patient with congenital tricuspid stenosis who was treated successfully by a palliative open operation. The preoperative diagnosis was tricuspid atresia. At operation, however, congenital tricuspid stenosis was detected and managed by commissurotomy.. The postoperative course was very good. Later, the associated ventricular septal defect (VSD), pulmonary annular stenosis, and probably the atrial septal defect (ASD) will be corrected. We emphasize the importance of diagnosing congenital tricuspid stenosis correctly, because of the possibility of operating upon the valve itself.


A palliative operation for all types of aortic arch interruption in the neonate. A palliative operation is described for the treatment of infants with all types of aortic arch interruption without regard to anatomic variation. It consists of establishment of continuity between the main pulmonary artery and descending thoracic aorta with a prosthetic graft, pulmonary artery banding distal to the graft, and ligation of the closing ductus arteriosus. Infant mortality rate in the past has been high, rega-dless of the type of operation used. The method of treatment described in this report is one in which an initial palliative operation is employed in the newborn period with total correction planned at a later date, when the risk of morbidity and death will be less.


Periampullary carcinoma.

Periampullary carcinoma. In the majority of instances, periampullary tumors include adenocarcinomas of the pancreatic head, duodenum, ampulla of Vater, and lower bile duct. Diagnosis is based mainly on a history of jaundice or is made by endoscopic duodenoscopy with retrograde pancreatography or by cholangiography or both. The best treatment for these tumors is pancreatoduodenectomy or palliative bypass if the tumor has spread beyond the region encompassed by resection. In experienced hands, resection can be accomplished with a mortality rate of less than 10 per cent and is followed by a 5-year survival rate of 30 to 40 per cent in carcinomas of the ampulla, duodenum, or lower bile duct and of about 10 to 15 per cent in carcinomas of the pancreatic head. Palliative surgical, chemotherapeutic, and radiotherapeutic procedures as yet do not prolong life appreciably.


A multidisciplined approach for the treatment of metastatic carcinoma of the breast. We have reviewed our experience in a multidisciplined breast cancer clinic where we have utilized hormonal, ablative, and chemotherapetuci modalities. Our experience seesm to be similar to that of other groups in that oophorectomy treatment produces approximately a 61 per cent response (regression and arrest) rate, androgen therapy produces a 47 per cent response (regression and arrest) rate estrogen therapy produces a 40 per cent response (regression and arrest) rate, and ablative treatment produces approximately a 50 per cent response (regression and arrest) rate. Adrenalectomy and hypophysectomy showed similar response rates. Until it can be shown that hypophysectomy clearly offers enhanced benefits, this will not be utilized by our group except in those patients who cannot tolerate abdominal surgery (that is, patients with poor pulmonary reserve). Of interest is the high response rate (65 per cent) to ablative treatment in patients in whom disease exacerbates on additive hormonal treatment, with an increased duration of response and survival. Survival is increased in patients who are rebound responders after estrogen withdrawal. We expect to report data with future follow-up of this group of patients. New protocols will be instituted after review of the data in the hope of increasing clinical benefit and survival in this group of patients. Carcinoma of the breast accounts for almost 90,000 new cases of cancer a year, with metastases eventually developing in at least half of these patients. All physicians must be aware of the many complex problems associated with this disease and, hopefully, arrive at a logical approach for its control. We believe this can be achieved with a multidisciplined group approach as established at the Lahey Clinic Foundation.


Control of ventricular arrhythmias during myocardial infarction by antilipolytic treatment using a nicotinic-acid analogue. The effect of lowering raised plasma-free-fatty acids (F.F.A.) on the incidence of serious ventricular arrhythmias after myocardial infarction was assessed by a double-blind trial in eighty-one patients. A nicotinic-acid analogue (N.A.A.) with very slight haemodynamic effects was given within 12 hours of the onset of myocardial infarction to lower plasma-F.F.A. When treatment with N.A.A. was started within 5 hours of the onset of symptoms, the numbers of patients with ventricular symptoms, the numbers of patients with ventricular tachycardia were significantly reduced, provided elevated plasma-F.F.A. levels were rapidly lowered and maintained in the normal range throughout the treatment period. The incidence of R-on-apex T ventricular premature beats and beats in which the ectopic R wave interrupted the apex of the T wave of a previous ventricular premature beat was also reduced in patients receiving N.A.A within 5 hours of the onset of symptoms. Plasma-total-catecholamines and serum-creatine-kinase levels were similar in the N.A.A.-treated and placebo groups. N.A.A. rarely caused skin flushing, but vomiting occurred in some patients after many hours of treatment. These findings suggest that treatment directed towards stabilsing the matabolism of the ischaemic myocardium can be of therapeutic value and lead to fewer serious ventricular arrhythmias.


A virological and epidemiological study of patients with acute hearing loss. Virological and epidemiological data were examined in thirty-nine patients with acute hearing loss.  The hearing loss was unilateral in thirty-one patients and bilateral in eight.  There was a history of upper-respiratory-tract infection before onset of deafness in ten of the patients.  Cases were more common in the autumn, with nineteen of the cases occurring in the 3 months from September to November.  About half of the patients were 20-40 years old and half 46-73 years old; only one patient was aged less than 20 years.  Antibodies to several common viruses were titrated, and there was evidence of recent infection by mumps virus in one case and by Mycoplasma pneumoniae in another.  Although no evidence of recent virus infection was found in the remaining patients, it is suggested that the condition may be viral in origin and that further studies should be undertaken.


Amines, anticonvulsants, and epilepsy.

Amines, anticonvulsants, and epilepsy. Concentrations of 5-hydroxyindoleacetic acid (5-H.I.A.T.) in cerebrospinal fluid (C.S.F.) were significantly raised in twenty-seven anticonvulsant-treated epileptic patients compared with fifteen untreated epileptics and twenty-two neurological controls. This rise was not seen until therapeutic blood-levels of phenobarbitone and diphenylhydantoin had been achieved, and was most striking in clinically intoxicated patients. Similar trends were seen in C.S.F. homovanillic acid (H.V.A). There was a close correlation between C.S.F. 5-H.I.A.A. and H.V.A., especially in the treated epileptics. These findings have implications for the antiepileptic and toxic effects of anticonvulsant drugs.


Urine microscopy as an aid to detection of bacteriuria. Fresh, uncentrifuged urine specimens obtained from children attending a renal clinic were examined microscopically for leucocytes and bacteria, and the results were compared with quantitative bacterial cultures.  Of the 51 specimens giving colony-counts larger than or equal to 10-5 per ml., 6 (12 per cent) showed no microscopical bacteriuria and 22 (43 per cent) contained smaller than 10 leucocytes per c.mm.; only 3 specimens (6 per cent) showed neither bacteria nor excess cells.  Of the 186 uninfected specimens, bacteria were seen microscopically in 32 (17 per cent) and an excess of leucocytes in 27 (15 per cent); in only 9 (5 per cent) were both observed.  Leucocyte-counting alone is therefore an unreliable technique, but microscopy for both bacteria and leucocytes affords a simple and rapid method of preselecting urine specimens for culture, with a high probability of predicting the correct result.  While not suitable for population screening, it is ideal for use in hospital clinics and wards, as well as in general practice.


Nicotine absorption by workers harvesting green tobacco. Green-tobacco sickness is an occupational illness of tobacco harvesters.  Symptoms include nausea, vomiting, dizziness, and prostration.  The disease is self-limited and of short duration, but recurs frequently in susceptible workers.  The aetiology is not known, but nicotine has been suspected as a causative agent.  Thirty-two workers on four North Carolina tobacco farms were studied during harvesting.  None of these workers smoked or chewed tobacco.  Urinary cotinine (the major metabolite of nicotine) levels were monitored over a 24-hour period to evaluate nicotine absorption.  There was a tenfold rise in mean excretion of cotinine among workers who had greatest contact with the tobacco.  Less cotinine was found in urine of workers who had less exposure.  Levels of cotinine exceeded those found in novice smokers who smoked 3 cigarettes in succession.  Absorption of nicotine from tobacco leaf is the likely cause of tobacco sickness.


Assessment of outcome after severe brain damage. Persisting disability after brain damage usually comprises both mental and physical handicap.  The mental component is often the more important in contributing to overall social disability.  Lack of an objective scale leads to vague and over-optimistic estimates of outcome, which obscure the ultimate results of early management.  A five-point scale is described--death, persistent vegetative state, severe disability, moderate disability, and good recovery.  Duration as well as intensity of disability should be included in an index of ill-health; this applies particularly after head injury, because many disabled survivors are young.


Gallbladder disease in hyperlipoproteinaemia. The occurrence of gallbladder disease (G.B.D.) (cholelithiasis, cholecystitis, cholecystectomy) was examined in patients consecutively admitted beccause of hyperlipoproteinaemia types IIa and IV. Altogether 37 of the 52 patients with the type IIa pattern were women, whereas 56 of the 75 subjects with hyperlipoproteinaemia type IV were men. The overall incidence of G.B.D. in the group with the type IIa was 13 per cent in the males and 22 per cent in the females; the corresponding figures in type IV were 41 per cent and 68 per cent, respectively. The findings in the major age-group (40-59 years) were compared with those from three necropsy series covering subjects of the same age. The incidence of G.B.D. was then found to be normal in type IIa but abnormally high in type IV. Patients with and without G.B.D. did not differ with regard to body-weight or glucose tolerance.


A controlled trial of intra-articular radiocolloids versus surgical synovectomy in persistent synovitis. The results of a randomised trial of irradiation of the knee (synoviorthése), by intra-articular injection of yttrium-90, and surgical synovectomy have been compared in twenty knees in seventeen patients.  The mean length of follow-up was 2 years.  Relapse occurred in three out of ten irradiated knees, and in 2 out of 10 operated knees.  Fewer irradiated knees were involved when generalised exacerbations of polyarthritis occurred.  Irradiation carries a lower risk of complications than does synovectomy, it is more acceptable to patients, requires fewer days in hospital, and is cheaper; it would seem to be the treatment of choice in the older patient.


Diuretic-induced oedema.

Diuretic-induced oedema. Diuretics are now commonly prescribed for women with so-called idiopathic oedema.  Two patients with idiopathic oedema, one of whom was oedematous on diuretics, had the diuretics stopped.  In one the oedema became much worse, and in the other oedema developed.  However, this oedema disappeared spontaneously in both patients and they have remained free of oedema on no treatment.  It is suggested that in some patients treated with diuretics compensatory mechanisms to retain sodium and water may be stronger than the diuretic-effect itself and may lead to perpetuation of unnecessary treatment.


Simple method for automating the differential leucocyte-count. A simple method is described for analysing leucocyte-volume distribution curves to obtain a differential leucocyte-count.  The results derived by this method agree well with the differential counts obtained by microscopical examination of the stained blood-film.


Reaction to Kveim test material in sarcoidosis and other diseases. 81 patients, 24 with sarcoidosis and the remainder with various other diseases, were tested with three batches of Kveim material prepared from the same spleen. Positive Kveim tests were observed in sarcoidosis, tuberculosis, Hodgkin's disease, ulcerative colitis, rheumatoid arthritis, and Weber-Christian disease. These results show that a positive Kveim reaction to the material under test was not specific to sarcoidosis.


Absence of effect of bran on blood-lipids. Healthy medical students were divided into two groups.  One group was given double the normal fibre intake of their diet as a high-fibre-containing breakfast cereal; the other group (control) was given a breakfast cereal containing negligible amounts of dietary fibre.  Otherwise the nutrient intake of the two groups was maintained constant.  No differences in the serum-cholesterol, serum-triglyceride, serum-calcium levels were found between the two groups during or at the end of the study.  The addition of a tolerable amount of dietary fibre to the diet does not affect serum-lipids.


Faecal bile-acids and clostridia in patients with cancer of the large bowel. Of 44  patients with cancer of the large bowel, 36 ( 82%) had high faecal bile-acid concentrations compared with only 15 (17%) out of 90 patients with other diseases. 31 (70%) of the 44 patients with large-bowel cancer had high faecal bile-acid concentrations in the presence of faecal clostridia able to dehydrogenate the bile-acid nucleus, compared with only 8 (9%) out of 90 patients with other diseases. Thes findings support the hypothesis that cancer of the large bowel is caused by high concentrations of bile-acid derivatives produced by certain anaerobic bacteria.


Relation between regular intake of phenacetin-containing analgesics and laboratory evidence for urorenal disorders in a working female population of Switzerland. A study group of 623 employed Swiss women aged 30-49 years showing objective evidence of intake of phenacetin-containing analgesics, and a control group of 621 comparable women showing no such intake, were observed for 4 years (1969-72) for laboratory evidence of urorenal disorders. In both study and control groups morbidity was low. There was no difference between the study and control groups with respect to subsequent proteinuria, bacteriuria, and haematuria. The 4-year incidence of low urine specific gravity after overhight thirsting was significantly higher in the study group than in the control group (3-8% v. 0-8%) and the incidence of raised serum-creatinine was also significantly higher in the study group (2-9% v. 0-4%). However, when the study group was further subdivided into a sub-group showing evidence of high intake of phenacetincontaining analgesics and one showing low intake, only the high-intake subgroup had an incidence of raised serum-creatinine (5-4%) significantly higher than the control group (0-4%), whereas the low-intake subgroup had an incidence (0-4%) similar to the control group.


Migraine and reporoductive hormones throughout the menstrual cycle. In a clinical survey the relation between migraine and menstruation was studied in 142 otherwise healthy women. In 24, onset of migraine coincided with the year of menarch. Of the 138 patients in whom onset of migraine predated the menopause, there were only 13 in whom attacks occurred regularly, and only, just before or during menstruation; in a further 11 attacks occurred regularly in relation to menstruation and at other times. Those with menstrually related migraine were more likely to have onset of migraine at menarche, to have associated weight gain and breast discomfort as part of a periodic syndrome, and to show improvement during pregnancy. There appeared no clear pattern of change at the menopause. In a study of reproductive hormones, blood was collected daily throughout a menstrual cycle from each of 8 women with menstrually related migraine, 6 with menstrually non-related migraine, and 8 healthy headache-free controls. Plasma levels of follicle-stimulating hormone (F.S.H.), luteinising hormone (L.H.), prolactin, oestrogen, and progesterone were measured in all. Plasma-testosterone was measured in 8 migraine patients. Mean plasma oestrogen and progesterone levels were significantly higher in migraine patients than controls for most of the menstrual cycle, with the most striking differences found in the late luteal phase for progesterone. No significant difference was found between the menstrually related and non-related patients for these or the other hormones measured. Mean plasma-prolactin levels were lower in migraine subjects than controls, but the difference was not significant. Mean plasma F.S.H. and L.H. levels were similar in both migraine and control groups. Plasma-testosterone levels were within the range for normal in the 8 migraine patients studied. No specific hormone changes were associated with the occurrence of a migraine attack, nor did rising or falling levels, or greater increments of change over given cycle phases, appear important in provoking attacks.


Is low-renin hypertension a stage in the development of essential hypertension or a diagnostic entity? A study of the frequency distribution of plasma-renin concentration in 81 patients with essential hypertension produced no evidence of a distinct sub-population with low renin levels. An arbitrary dividing line was used, therefore, to define low-renin hypertension (36% of patinets). Patients in this group were older than those with normal renin levels, and there was a significant negative correlation between renin and age among all patients. Low-renin hypertension was not characterized by increased exchangeable sodium, but exchaneable postassium was significantly lower than in patients with normal plasma-renin. This difference became insignificant when five patients in the low-renin group with persistent hypokalaemia were excluded. It is concluded that low-renin hypertension does not represent a separate diagnostic entity but that plasma-renin falls with age in essential hypertension.


Chronic pancreatitis, sclerosing cholangitis, and sicca complex in two siblings. A new syndrome of chronic pancreatitis, sclerosing cholangitis, sicca complex, and other features was found in a brother and sister. Leucocyte-migration inhibition in the presence of a bile antigen in both patients suggested that immune mechanisms may be involved.


Vitamins and oral contraceptive use.

Vitamins and oral contraceptive use. Reports concerning the interaction between steroidal contraceptives (the combined pill) and vitamins indicate that in users the mean serum-vitamin-A level is raised and the mean serum-vitamin-B2 (riboflavine), vitamin-B6 (pyridoxine), vitamine-C, folic-acid, and vitamin-B12 levels are reduced. Other vitamins have been insufficiently studied for comment. Biochemical evidence of co-enzyme deficiency has been reported for vitamin B2, vitamin B6, and folic acid. Clinical effects due to vitamin deficiency have been described for vitamin B6--namely, depression and impaired glucose tolerance. Folic-acid deficiency with megaloblastic anaemia has been reported in only 21 cases.


Effusions into body cavities in hypothyroidism. Effusions into body cavities secondary to hypothyroidism, though rare, are well recognised. They may precede the more specific and typical manifestations of the disease, thereby misleading the clinician.


Immediate prognosis in recurrent myocardial infarction. Of 880 patients admitted consecutively to a coronary-care unit with acute myocardial infarction, 200 had had a previous infarction.  The mortality-rate in the first twenty-eight days after infarction was higher in the group with recurrent infarction (26 per cent) than in the group with first infarctions (16.9 per cent).  When the patients were matched for sex and age this difference in mortality-rate was only statistically significant in men aged less than seventy.  In patients with previous infarctions the short-term prognosis was worse in those with two or more previous infarctions, in those in whom infarction had occurred less than three months before, and in those with a previous infarction in the anteroseptal wall.


Hazards from simian herpes viruses: reactivation of skin lesions with virus shedding. A new simian herpes virus with biological properties similar to herpes simplex and to simian "B" virus has been used as a model system for studying virus latency in dorsal root spinal sensory ganglia.  Following intradermal injection, virus is present in the skin lesions and corresponding ganglia only, during the acute stage of the disease.  By organ-culture techniques, latent virus was rescued from ganglia up to 2 years later.  No latent virus was ever found in skin organ cultures of the primary site.  Treatment with cortisone up to 18 months later reactivated virus latent in the ganglia, and virus returned to the skin where it produced small but typical herpes lesions which shed virus.  Reactivation of Herpesvirus tamarinus was achieved after 28 months.  This is believed to be the first report of a model system for the study of herpes latency in which skin lesions are found to recur, and provides an opportunity for more detailed investigations of the mechanisms of virus latency in man.  The presumption that reactivation of skin lesions will also be possible in rhesus monkeys seropositive for "B" virus points to a possibly grave and largely unsuspected hazard for those engaged in primate research.


Passive transfer of antibodies of maternal origin from blood to cerebrospinal fluid in infants. The demonstration that specific IgM antibodies are present in the serum of infants is useful in the diagnosis of several congenital infections.  However, it is less certain whether the detection of antibodies in cerebrospinal fluid (c.s.f.) of infants indicates congenital infection of the central nervous system, because the origins of such antibodies have not been established.  In the present study diphtheria and tetanus antitoxins of maternal origin have been detected both in the serum and in the c.s.f. of infants.  These observations suggest that an important source of immunoglobulins in c.s.f. is passive transfer of antibodies from serum which should be considered in interpreting serological studies with c.s.f.


Refeeding-malaria and hyperferraemia.

Refeeding-malaria and hyperferraemia. During the Central African (Sahelian) drought, attacks of falciparum malaria were common in patients and their relatives shortly after their arrival in a hospital in Eastern Niger.  A prospective study of 72 adult patients not admitted for malaria and 109 accompanying relatives was undertaken to investigate this observation.  23 attacks occurred in patients and 51 in relatives, with a peak frequency five days after arrival.  On arrival, parasitaemia was low but reached a maximum by five days.  Serum-iron and percentage saturation of transferrin were moderately increased initially, rose dramatically within forty-eight hours with near maximum saturation, and were falling by the fifth day.  It is suggested that the early hyperferraemia, apparently related to refeeding, led to rapid multiplication of existing parasites and attacks of malaria.  The results of experimental malarial infection of Wistar rats, half of which had been given intramuscular iron, supported this hypothesis.


Bottle-feeding: influence of teat-hole on suck volume. Infants have been fed with teats whose end-holes varied in shape and size. Two types of feeding-bottles were used. With conventional feeding-bottles, variations of teat-hole shape and size did not significantly alter the volumes of milk obtained at each suck. However, when an infant feeding-bottle was used which contained a valve, then variations in teat-hole shape and size did alter the volumes of milk obtained. The volume of milk obtained from a standard teat under such circumstances was the same as the volume previously calculated for breast-feeding.


Pyogenic liver abscess caused by Streptococcus milleri. Three clinically typical cases of multiple pyogenic liver abscesses are presented.  In all three cases Streptococus milleri was isolated from the pus and in two cases it was isolated from blood cultures also.  The patients were successfully treated by surgical drainage and antibiotics.


Immunogenicity and acceptability of a human diploid-cell culture rabies vaccine in volunteers. A rabies vaccine, prepared in human diploid-cell culture, was given intradermally (i.d.) or intramuscularly (i.m.) to 35 volumteers who had not previously received prophylactic immunisation.  Rabies neutralising antibody titres rose sharply after the first dose, and 4 weeks after the second dose all titres were above 1/80 (1-7 i.u. per ml.).  Immunisation via the i.d. and i.m. routes was equally successful in terms of likely immune status, but the i.d. rate was associated with many more local sideeffects.


Disorders of blood-lipids in renal disease. Hyperlipidaemia is a characteristic feature not only of the nephrotic syndrome but also of chronic renal disease without the features of that syndrome.  There is evidence for disordered lipid metabolism in patients with chronic renal disease.  In these patients the disordered lipid metabolism, the precise cause of which is unknown, is characterised by hypertriglyceridaemia, the aetiology of which is probably multifactorial.  Hyperlipidaemia is an important potential risk factor in the aetiology of cardiovascular disease, which may be a leading cause of death in patients undergoing long-term maintenance haemodialysis therapy.


Costs of detecting and treating cancer of the uterine cervix in North-East Scotland in 1971. In 1971 the Aberdeen cytology service handled 22,291 cervical smears--threequarters from women participating in a screening programme and the rest from women with symptoms who were referred to hospital.  As a result of screening, 56 preclinical cases of cancer of the cervix uteri were treated in hospital; 13 others were classified as less than preclinical.  Of the gynaecological patients 20 had clinical cancer and 29 had less than preclinical lesions.  From estimates of the costs of running the cytology service and of hospital inpatient costs it is possible to derive figures for detecting and treating preclinical cases and investigating and treating clinical cases.  The cost to the Health Service of detecting and treating each preclinical case was slightly less (445 pounds) than that for impatient treatment of each clinical case (487 pounds).  However, if mass screening were abandoned cytology would almost certainly continue for women referred to hospital with symptoms, and, if the costs of taking and examining these smears is taken into account, the cost per clinical case treated nearly doubles to 835 pounds.  Had outpatient and other follow-up costs been included, the difference in service costs would be even greater.  Comparison of these figures assumes the controversial point that preclinical cancer will always progress to invasive carcinoma if left untreated and takes no account of inflation and discounting.


[Characteristics of artefacts following fixation with glutaraldehyde and osmium tetroxide]. In the myocard and kidney of rats perfused with Karnovsyk's solution for 5--10 minutes then fixed for two more hours and postfixed in 1 per cent solution of osmium tetroxid in Millonig buffer, characteristic electrondense granules were observed. The structure of the organ appeared to be normal. Although the granules appeared to be diffuse, they presence was most marked on the membranes, on the surface of erythrocytes and in the pinocytic vacuoles both on counterstained slides and on those of without counterstaining. When postfixation was carried out with osmium-tetroxide solved in Veronal buffer artefacts did not occur. To avoid artefacts fixation in glutaraldehyd should not be followed by a postfixative containing phosphate buffer.


[Experimental malakoplakia in the kidney]. Malakoplakia in kidney of rats was caused experimentally by injection of large quantities of not purified endotoxin-antigen complex of Escherichia coli 0 75 (O. coli 127 97 CDC 0 group 75; American Type Culture Collection, Eight Edition 1968, page 24). The extract in the beginning became surrounded by leukocytes, later by macrophages. The latter then were transformed into characteristic Hansemann cells. From the 8th day on started the depletion of calcium phosphate in the cytosegresomes of macrophages, thus forming the Michaelis-Guttmann bodies, necessary for the diagnosis of malakoplakia. It is believed that complex of endoxin- and antigen of Escherichia coli may take a part - at least in certain cases - in formation of malakoplakia in human.


[Electron miscroscopic liver changes in acute carbon tetrachloride poisoning in man)]. Lesions of the liver revealed by light and electron microscopy at two young men, workers of the chemical industry are described. At one of the patients biopsy was performed twice at the other three times. Patient having jaundice were admitted to hospital because of the suspected acute viral hepatitis. Laboratory findings have shown an acute parenchymal lesion. By light microscopy at one of them mid-zonal, at the other centro-lobular necrosis of hepatic cells further lipoid droplets of the cytoplasma of hepatocytes were revealed. Electronmicroscopically in the first biopsy specimens, and at one of the ptients at the second biopsy as well mostly damage to the mitochondria (enormous osmiophilia of the matrix, presence of osmiophil granules) and enlargement of the tubules of the endoplasmic reticulum could be seen. The controll clinical and morphological findings evidenced the reversible character of the process. Authors stress the differences existing between hepatic lesions caused by CCl4 at different persons on one side, and at rat and human liver on the other.


Induction of translocations by cyclophosphamide in different germ cell stages of male mice: cytological characterization and transmission. Cytological and fertility tests were performed in F1 male mice derived from different germ-cell stages of male parents treated with cyclophosphamide (350 mg/kg body weight). The objectives of the present experiment were: (I) to determine the sensitivity of the male germ-cell stages to the induction of translocations by the compound, and (2) to characterize translocation configurations in F1 and F2 males, in order to obtain information about the pattern of chromosome breakage induced and its transmission to subsequent generations. Of 508 F1 males studied, 39 were partially sterile. The group of males conceived 8-21 days after treatment contained by far the highest proportion of partially sterile animals (30%). It was also the only group in which totally sterile animals (11%) were found. Of 25 semisterile males from this group, 24 gave evidence of translocations when spermatocytes were scored at diakinesis. The translocation frequencies in F1 derived from treated spermatozoa and spermatocytes were 14 and 1%, respectively. No translocations were detected cytologically in 6 semisterile males derived from treated spermatogonial stages. These results indicate that spermatid stages are especially sensitive to the mutagenic action of cyclophosphamide. In 21 of the 31 semisterile translocation males (68%), the majority of the spermatocytes contained 18 bivalents plus a ring-of-four configuration, indicating that both breakpoints were relatively centrally located; and in several of these males, the frequency of cells with rings was close to 100%. In another 9 F1 males (29%) the predominant multivalent configuration was a chain-of-four, indicating one of the breakpoints to be relatively more terminally located; and in one male (3%), the majority of cells had two unequal bivalents, indicating both breakpoints to be fairly close to the ends of the chromosomes involved. Determination of centromere positions by the use of C-banding showed that chain-of-four configurations in any one male were predominantly of a given type..


Activation of the properdin pathway of complement in patients with gram-negative of bacteremia. To determine the pathway used for activation of complement component C3, serum levels of components C1, C4, C2, C3, C5, C6, and C9 and two properdin factors, properdin and factor B, were measured in 42 patients with gram-negative bacteremia, in 19 of whom shock subsequently developed. Mean levels of the classical components C1, C4, and C2 in bacteremic patients in whom shock subsequently developed did not differ significantly (p greater than 0.05) from those of patients with uncomplicated bacteremia. Levels of properdin, factor B and C3, C5, C6, and C9 were significantly (p less than 0.05) decreased in patients with shock in comparison with those with uncomplicated bacteremia. Taken together, these findings are consistent with activation of C3 and the terminal complement sequence, C5-C9, occurring primarily by the properdin pathway, in patients with gram-negative bacteremia eventuating in shock. Biologically active products released during activation of C3-C9 may contribute to the development of shock.


Chronic progressive panencephalitis due to rubella virus simulating subacute sclerosing panencephalitis. Late-onset chronic progressive panencephalitis developed in a 12-year-old boy with congenital rubella syndrome from whose brain rubella virus was isolated. Progressive dementia began at eight, and ataxia, choreiform movements, myoclonic seizures, and fine perimacular pigmentation appeared at 11 years of age. The cerebrospinal fluid was minimally pleocytotic and had a total protein of 156 mg per deciliter, of which 52 per cent was gamma globulin. Electroencephalography demonstrated generalized medium and occasional high-voltage slowing without burst suppression. The antibody titer to rubella virus (hemagglutination inhibition) was 1:8192 in serum and 1:256 in cerebrospinal fluid. Antibody titer to measles virus (complement fixation) was less than 1:8 in serum. Microscopical study of biopsied brain tissue at the age of 11 disclosed panencephalitis similar to subacute sclerosing panencephalitis, but with perivascular deposits and without inclusion bodies. Rubella virus was isolated from the brain by cocultivation with CV-1 cells.


[Eosinophilic gastroenteritis. Report of a clinical case]. An unusual case of eosinophilic gastroenteritis is described, together with its differential diagnosis from allergic enteropathy. Numerous biopsies and other instrumental examinations pointed to small intestine as the only site and eosinophilic enteritis as a suitable classification. Disappearance of the clinical symptoms was obtained with corticosteroid management in the space of 6 months.


Progress in fetal assessment.

Progress in fetal assessment. The growth areas in fetal assessment in late pregnancy have been studied by making a quantitative review of the papers in four obstetric journals. Of the 130 relevant papers published, in 1973, the most common subject treated was the phospholipid test of fetal lung maturity. Other important tests reviewed were estrogen assays, ultrasonic studies, human placental lactogen and alpha-fetoprotein measurements. The clinical value of phospholipid tests was demonstrated; liquor creatinine assays are much inferior in assessing fetal maturity. Plasma estriol assays are likely to be of increasing importance in clinical practice. Some caution should be applied in the interpretation of ultrasonic measurements of biparietal diameter as a test of fetal development and well-being.


Some etiologic and prognostic factors in early infantile autism and psychosis. Fifty infants and young preschool children seen in a pediatric developmental service and diagnosed as having "autism" all had evidence of organic disease of the brain and three fourths had mental deficiency of varying degrees. They did not differ in any respect from a comparison group of patients with central nervous system dysfunction unassociated with the symptom complex of autism. Both groups of patients had a high incidence of low birthweight, complications of pregnancy and the neonatal period, seizure disorders, and a variety of specific disease entities associated with developmental defects. Follow-up of 40 of the 45 survivors for a mean of five years showed that none of the patients had had treatment directed to their psychotic symptoms. However, three fourths had established social responses appropriate to their level of function; those who did not generally were over 3 years of age at the time of their first examination or had initial DQs of 35 or less. The degree of mental deficiency was as great or greater at follow-up than it was initially.


Personality profiles and leadership potential of medical-surgical and psychiatric nursing graduate students. When personality profiles and leadership potential of psychiatric and medical-surgical nursing graduate students were compared, using the California Psychological inventory (CPI) and the Managerial Key for the CPI, medical-surgical majors scored significantly higher on the CPI scale (p greater than .05). Overall, both groups scored higher than the norm and showed a more optimal personality development than has been observed in earlier studies of this kind. In the Managerial Key for the CPI, the test of leadership potential on both groups received a high rating, comparable to that of the top-management group in the original research.


Differences in the surface proteins of mouse B and T cells. We have selectively labeled the surface of mouse thymus-dependent (T) and thymus-independent (B) lymphocytes by means of lactoperoxidase-catalyzed iodination. Examination of the labeled proteins by electrophoresis in the presence of sodium dodecylsulfate followed by autoradiography revealed differences in the cell surface proteins of B and T cells. Proteins from labeled thymocytes, T cell lymphomas, and both normal and activated peripheral T cells give a broad band of radioactivity corresponding to at least two protein components with apparent molecular weights of 170,000 to 190,000. This band is absent from autoradiographs of iodinated B cell proteins, which instead have another band corresponding to a protein with an apparent molecular weight of about 220,000. We have shown that these T and B cell marker proteins are synthesized by the cell and are not serum components selectively bound to the cell surface. We have also established that these proteins are the major iodinated species precipitated from Nonidet P-40 cell lysates by rabbit anti-mouse lymphocyte serum, suggesting that they are the major antigens recognized by rabbit antibodies to mouse lymphocytes.


Inhibition of oncornavirus functions by 2'-azido polynucleotides. The 2'-azido analogs of poly(U) and poly(C), poly(dUz) [poly(2'-azido-2'-deoxyuridylic acid)], and poly-(dCz [poly(2'-azido-2'-deoxycytidylic acid)], were found to inhibit the RNA-directed DNA polymerase (reverse transcriptase) activity of murine leukemia (Moloney, Rauscher) and sarcoma (Moloney) virus, and feline leukemia (Theilen) and sarcoma (Gardner) virus, while under the same conditions the unsubstituted parent compounds failed to do so. In addition, poly(dUz) and poly(dCz) inhibited the replication of exogenous murine sarcoma virus (Moloney) in nontransformed cells (as assessed by an infectious center assay), but poly(dUz) failed to suppress the formation of endogenous sarcoma and leukemia viruses in transformed cell lines (MO-P, JLSV5). In these same cells, poly(dUz) failed to inhibit the multiplication of vesicular stomatitis virus. These data add further strength to the contention that reverse transcriptase is necessary for the productive infection and transformation of normal cells by oncornaviruses but is not essential maintenance of this transformed state and the continuous production of new viruses particles by these transformed cells.


Enzymatic activation and trapping of luminol-substituted peptides and proteins. A possible means of amplifying the cytotoxicity of anti-tumor antibodies. Glutathione and glucose oxidase (EC 1.1.3.4) conjugates containing covalently bound luminol were prepared as prototypes for peptides and proteins with latent, enzyme-activatable chemical reactivity. In the presence of small quantities of activated horseradish peroxidase, conjugated luminol molecules were oxidized to unstable free radicals which reacted rapidly with soluble proteins and cells. These observations are of interest in regard to possible sequential localization reactions in which a few molecules of cell-bound antibody-horseradish peroxidase would be used to catalytically alter and trap many molecules of a second (luminol-substituted) enzyme, toxin, or hapten in the same area, as might be desirable in promoting selective cell destruction.


Site-directed mutagenesis: effect of an extracistronic mutation  on the in vitro propagation of bacteriophage Qbeta RNA. It has been proposed that the nucleotide sequences of the 3' terminal extracistronic regions of phage RNA plus and minus strands have been strictly conserved during evolution because they are stringently required for recognition by the viral replicase. We have devised a method to generate point mutations at selected sites in the genome of phage Qbeta. The in vitro synthesis of Qbeta RAN with G leads to A transition in the 16th position from the 3' end, i.e., in the terminal extracistronic region of the genome, is outlined. When a mixture of about 60% wild-type and 40% mutant RNA was repeatedly replicated, the mutant RNA was enriched to 80%, showing that at least this point mutation in the terminal sequence of Qbeta RNA does not impair its in vitro replication, but in fact slightly accelerates it.


Radiation therapy in multiple myeloma.

Radiation therapy in multiple myeloma. In this retrospective study of 76 patients with myeloma, the indications for radiation therapy are reviewed and its value analyzed. Eleven patients presented with a "solitary plasmacytoma" and 65 patients presented with disseminated disease. Radiation therapy is successful in both the palliation of pain (81% of cases) and in producing long-term remission and possibly cure in solitary lesions.


P-Chloramphetamine: Selective neurotoxic action in brain. Injection of 2.5,5, 10, or 20 milligrams of p-chloroamphetamine per kilogram of body weight into rats produced evidence of cytopathological changes in sections of brain stained by a Nissl or silver method. As early as 1 day after drug injection cells demonstrated an intense Nissl staining, intense argyrophilia, cellular shrinkage, and perineuronal spaces. At 30 days after injection both stains revealed cellular debris and glial reactions characteristic of cellular dissolution. The neurotoxic effects of 2.5, 5, or 10 milligrams of p-chloroamphetamine per kilogram were primarily restricted to an area of the ventral midbrain tegmentum corresponding to the distribution of the B-9 serotonergic cell group. After 20 milligrams of p-chloroamphetamine per kilogram there was also evidence of neurotoxic effects on cells within the substantia nigra. These results confirm previous suggestions that the long-term reduction in serotonin content of brain, tryptophan-5-hydroxylase activity, and uptake of serotonin after injection of p-chloroamphetamine is due to a neurotoxic effect of the drug or some metabolite on serotonergic cell bodies.


Mechanism of axonal transport: a proposed role for calcium ions. In vitro axonal transport of tritiated protein decreased 40 to 60 percent when neuronal cell bodies were incubated in calcium-free medium, but was not affected when only nerve trunks were exposed to calcium-free conditions. In addition, calcium-45 was transported along axons at a rate similar to that of rapidly transported tritiated protein. These data are interpreted to suggest that calcium ions are involved in the initiation of axonal transport and in the coupling of transported proteins to the transport system.