Prostaglandins and asthma: The use of blood components for metabolic studies. Experiments were carried out to study a possible role of prostaglandins (PGs) in the pathogenesis of human bronchial asthma. Leukocyte, plasma, and serum of ambulatory asthmatic as well as nonasthmatic individuals were used. PGF was measured by competitive radioimmunoassay with anti-PGF2alpha antibody. It was found that PGF was spontaneously released from the leukocytes of both asthmatic and nonasthmatic individuals. The PGF release as studied with the cells of asthmatic individuals was enhanced by the additions of arachidonic acid and/or dibutyryl cyclic AMP. Antigenic challenge of sensitive cells failed to show any general effect on the amount of PGF released. The amount of spontaneously released PGF had no correlation with the amount of maximal antigenic histamine release. Preincubation of the cells with arachidonic acid had no effect on subsequent antigenic histamine release. Plasma PGF levels in nonasthmatic and in asthmatic individuals were not significantly different and both were considerably lower than the serum levels. The mean serum PGF level in the asthmatic group was higher than in the nonasthmatic, indicating probably a greater release of PGF from blood cells.


Polymerized ragweed antigen E. IV. The analysis of the stability of molecular size and antigenic reactivity of the polymer. Ragweed antigen E polymerized by treatment with glutaraldehyde offers a potentially improved method for immunotherapy of human IgE-mediated pollinosis because studies have demonstrated that the polymer retains antigenic reactivity but has decreased ability to release histamine when these functions are compared on a weight basis. The potential clinical usefulness of the polymer is dependent both on stability of the polymer and on retention of antigenic reactivity. The method used for the analysis of the stability of the polymer under conditions of storage was the comparison of gel filtration fractionation profiles. No depolymerization was demonstrated during storage conditions for 2 mo, and only minimal evidence of additional polymerization (less than 8%) or depolymerization (less than 7%) was observed after 6 mo of storage. There was no difference in the per cent of polymerized antigen E precipitated by rabbit antiserum after storage as compared with control samples.


Differential cellular immune responsiveness to systems of the major histocompatibility complex. Antigens determined by genes of the major histocompatibility complex can be divided into two systems" the LD (lymphocyte-defined or L determinant) and SD (serologically defined or S determinant) antigens. This division is based on the differential cellular responsiveness to these two sets of antigens of two subpopulations of T lymphocytes: a proliferating helper cell responds primarily to the LD antigens while a cytotoxic T lymphocyte responds most strongly to the SD antigens.


An Index of Nutritional Quality for a balanced diet. New help for an old problem. The Index of Nutritional Quality (INQ) is a method of quantitative and qualitative analysis of single foods, meals, and diets which has special significance in assessing clinical nutritional problems. The INQ is a ratio of the nutrient-to-calorie content of foods which may be calculated by computer and printed as bar graphs and tabular data. The number of nutrients and the nutrient standards used for analysis are flexible parameters which may be varied for each clinical situation. Illustrative examples include INQ analysis of simple foods, an institutional house diet, the diabetic Exchange list, and the diagnostic evaluation of the dietary intake of a hospitalized patient.


Identification of a macrophage-specific cell surface antigen. A mouse-specific macrophage antigen (MSMA) was identified in NP-40 extracts of 125I-radiolabeled mouse preitoneal macrophages by using a rabbit anti-mouse macrophage serum (AMS) and SDS-polyacrylamide gel electrophoresis. The antigen was shown to have a m.w. of 83,000 daltons and was present on both normal and "activated" peritoneal macrophages. MSMA was also present on syngeneic adherent spleen cells, allogeneic peritoneal macrophages, a mouse macrophage cell line (P388D1), and exhibited some cross-reactivity with peritoneal macrophages from closely related species (rats and hamsters). MSMA was not present on nonadherent peritoneal exudate cells, spleen cells, erythrocytes, thymocytes, or bone marrow cells. Extensive absorptions of AMS with thymocytes and erytrocytes from mice were necessary to remove other antibodies that reacted with other mouse membrane antigens. An antiserum directed against a specific membrane antigen has great potential in elucidating structure-function relationships with regard to a number of macrophage activities.


In vitro immune response to the 2,4,6-trinitrophenyl determinant in aged C57BL/6J mice:changes in the humoral immune response to, avidity for the TNP determinant and responsiveness to LPS effect with aging. An in vitro anti-TNP response of the spleen cells from aged C57BL/6J mice showed approximately 4-fold less PFC than did that from young adult mice. Anti-theta serum-treated young spleen cells gave an anti-TNP response that was definitely greater than the response of the anti-theta serum-treated aged spleen cells in the presence of the exogenous activated thymus cells as helper cells. These results suggest that the deficits in B cells may be partly responsible for the imparied anti-TNP response of the aged spleen cells. To examine further the capacity of stem cells in the bone marrow to generate B cells responsible for anti-TNP response in the spleen, we injected i.v. 1.5 to 2.0 times 10(7) bone marrow cells from young or aged mice into lethally irradiated syngeneic recipients that had previously been thymectomized. Four to 6 weeks later, 10(7) spleen cells from the two groups of these recipient mice were immunized with TNP-SRBC in the presence of the exogenous activated thymus cells and assayed for anti-TNP PFC. The response of the aged marrow-derived B cells was approximately one-half of that of the young marrow-derived B cells. The avidity for TNP determinant of the antibodies produced by the PFC was determined by the plaque-inhibition technique. The avidity of the antibodies produced by the aged mice was approximately 33 times lower than that by the young mice. Anti-TNP response of the young spleen cells were markedly enhanced by the addition of LPS to the cultures, whereas no or little enhancement of the response was induced in the aged spleen cells even in the presence of high concentration of LPS. In contrast, DNA synthesis of both the young and aged spleen cells was comparably stimulated by 1 mug/ml and 10 mug/ml of LPS, however, it was rather less in the aged spleen cells at a concentration of 100 mug/ml. Mechanisms responsible for the changes in avidity and responsiveness to LPS with aging are discussed.


In vitro studies of the genetically determined unresponsiveness to thymus-independent antigens in CBA/N mice. The X-chromosome-linked B lymphocyte defect of CBA/N mice has been studied in vitro by comparing the ability of (CBA/N X DBA/2)F1 (X-/X- X X+/Y) male (X-/Y) and female (X-/X+) spleen cells to respond to the thymus-independent antigen DNP (or TNP)-AECM-Ficoll. (CBA/N X DBA/2)F1 male spleen cells failed to generate significant in vitro anti-TNP antibody responses to DNP- or TNP-AECM-Ficoll, in contrast to spleen cells from F1 female (X-/X+) mice which responded normally to these T-independent antigens. Spleen cells from male F1 mice responded almost as well as F1 female cells to the thymus-dependent antigen, TNP-sheep red blood cells (TNP-SRBC) in vitro. Adding F1 male cells to F1 female cells failed to reduce the response of the latter to DNP-AECM-Ficoll, suggesting that the inability of F1 male cells to respond was not due to active suppression. The response of F1 male spleen cells to TNP-SRBC was not impaired by adding high concentrations of TNP-AECM-Ficoll indicating that the mechanism of unresponsiveness was not tolerance induction in all TNP-specific precursors. Lymphocytes from F1 male mice were capable of forming anti-TNP antibody after stimulation with lipopolysaccharide (LPS) in high concentrations; DNP-AECM-Ficoll had no effect on this polyclonal response. B lymphocytes from mice bearing only the X-chromosome of the CBA/N strain thus display a profound defect in B cell activation. This functional defect may represent either an inability of the defective B cells to be activated by thymus-independent antigens or the absence of a sub-class of B cells which respond to thymus-independent antigens.


Immunosuppressive factor(s) extracted from lymphoid cells of nonresponder mice primed with L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT). The synthetic terpolymer of L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) not only fails to elicit a GAT-specific antibody response in nonresponder mice, but also prior injection of GAT specifically decreases the ability of nonresponder mice to develop a GAT-specific antibody response to a subsequent challenge with GAT-MBSA. This inhibition is mediated by GAT-specific suppressor T cells. Further, a suppressive factor can be extracted from lymphoid cells of GAT-primed nonresponder mice that inhibits the development of primary GAT-specific antibody responses to GAT-MBSA and to GAT-PRBC- by normal syngeneic mice. The suppressive activity is dose-dependent and absorbed by GAT-Sepharose, but not by BSA-Sepharose. The suppressive activity elutes from a G-100 Sephadex column in the same fraction as ovalbumin, suggesting its m.w. is approximately 45,000 daltons.


Lymphocyte transformation induced by autologous cells. III. Lymphoblast-induced lymphocyte to stimulation does not correlate with EB viral antigen expression or immunity. Human mitogen-induced and cell line B lymphoblasts stimulate the proliferation of allogeneic and autologous lymphocytes in culture. The role in thes reaction of EB viral determinants on the stimulating cells and immunity of the lymphocyte donor to the EB virus has been studied. The stimulatory capacity of cultured cell line lymphoblasts is not inhibited by incubating lymphoblasts with antisera to EB viral determinants. Cultured cell line B lymphoblasts stimulate as much thymidine incorporation by lymphocytes from donors with or without immunity to the EB virus. Further, a B lymphoblast cell line (U-698) which lacks the EB viral genome stimulated as much lymphocyte proliferation as did B lymphoblasts with the EB genome. Cultured T lymphoblast cell lines do not stimulate allogeneic lymphocyte proliferation. These cells appear to lack the determinants which stimulate lymphocyte transformation. No evidence was found that cultured cell line T lymphoblasts suppressed allogeneic lymphocyte proliferation. Mitogeninduced lymphoblasts from EB-immune and non-immune subjects stimulated the proliferation of autologous lymphocytes comparably. It is concluded that neither immunity to the EB virus nor expression of EB viral antigens on mitogen-induced on cell line lymphoblasts is necessary for the stimulation of lymphocyte proliferation.


Diversity of the antibody response to the different antigenic determinants on the hemagglutinin subunits of influenza viruses. Sera from rabbits hyperimmunized with hemagglutinin (HA) subunits isolated from the A/Port Chalmers/73 (H3N2)strain of influenza virus showed great differences in their cross-reactions with different strains of influenza virus. In hemagglutination-inhibition tests, some sera reacted to about the same titer with A/Port Chalmers/73 and A/Hong Kong/68 viruses, suggesting that these two strains were very closely related. Other sera, which reacted to high titer with A/Port Chalmers/73 virus, had only a low titer with the Hong Kong/68 strain, suggesting that the two viruses were distantly related. Evidence suggested that these diverse cross-reactions were due to widely different ratios, in the different sera, of antibodies to the "common" and the "specific" antigenic determinants on the HA subunits. Thus, some rabbits gave a stronger response to the "common" determinants than to the "specific", whereas in others, the reverse seemed to be the case. Sera from human volunteers injected with A/Port Chalmers/73 inactivated or subunit influenza virus vaccines, or from people infected with Port Chalmers/73 virus, contained, in most cases, antibodies predominantly to the "common" antigenic determinants on the HA subunits. These sera reacted to higher titer with Hong Kong/68 virus than with the Port Chalmers/73 strain. Absorption of these sera with Hong Kong/68 virus totally removed all detectable antibody, suggesting that they contained no antibody to the "specific" determinants of Port Chalmers/73 HA. Paradoxically, absorption of the sera with Port Chalmers virus did not remove all antibodies, suggesting that the sera contained antibodies to the "specific" determinants on Hong Kong/68 HA.


The effect of thiols on the immunologic release of slow reacting substance of anaphylaxis. II. Other in vitro and in vivo models. In the presence of L-cysteine, a selective and marked enhancement of the in vitro, immunologic release of slow reacting substance of anaphylaxis (SRS-A) from human peripheral leukocytes, sensitized monkey lung fragments, and sensitized guinea pig lung fragments was observed. In the rat, cysteine, but not sodium sulfide, enhanced the calcium ionophore (A23187)- induced release of SRS-A in vitro from mixed rat peritoneal cells and in vivo from the rat peritoneal cavity. Pretreatment of rats with cysteine also enhanced the IgGa-and anti-rat IgE-mediated release of SRS-A in vivo in the rat. These studies indicate a common biochemical mechanism involved in the formation and release of SRS-A from these different tissues and cells and further confirm the observation that the rat mast cell is not a major source of SRS-A in the rat.


Parital characterization of the rat anti-encephalitogenic protein (RSCP). A protein antigenically similar to the anti-encephalitogenic bovine spinal cord protein (BSCP) was detected in saline extracts of rat nervous tissues by immunodiffusion analyses using a rabbit anti-BSCP serum. Rat SCP (RSCP) appears to be evenly distributed throughout all parts of the rat nervous system and occurs also in the thymus, thyroid, and adrenal glands. Although immunodiffusion analyses indicated that RSCP shares some antigenic sites with BSCP, anti-RSCP sera reacted only with RSCP, indicating that the major immunogenic determinants of the RSCP are peculiar to the rat and differ from the immunogenic determinants of human, monkey, rabbit, guinea pig, or bovine SCP. Immunoelectrophoretic analyses of concentrated pastes of rat brain (RB) or rat spinal cord (RSC) in agar at pH 8.6 revealed that RSCP occurs in two molecular forms having the electrophoretic mobilities of a serum beta-globulin and a serum gamma-globulin, respectively. However, gamma-RSCP is the predominant component of extracts of brain or spinal cord. Gamma-RSCP was isolated from RB and RSC by a procedure which involved: a) extraction with 0.05 M ammonium acetate buffer, pH 4.0; b) batch absorption of impurities on CM-52 cellulose; c) batch absorption of RSCP on SP-Sephadex, pH 3.5; d) elution of RSCP from SP-Sephadex, pH 5.5; and finally, e) gel filtration on Sephadex G-50 superfine. Purified gamma-RSCP formed one band when analyzed by polyacrylamide electrophoresis in acid gels containing 8 M urea. In contrast, two bands were always present when gamma-RSCP from brain or spinal cord were subjected to SDS-polyacrylamide electrophoresis in 15% gels. The larger of the two components of brain gamma-RSCP had a m.w. of 12,400 daltons, whereas the two components of spinal cord gamma-RSCP were smaller. The molecular sizes of brain RSCP and spinal cord RSCP was estimated by gel filtration chromatography to be 12,400 daltons. The amino acid compositions of gamma-RSCP prepared from RB or RSC were similar except that gamma-RSCP from RSC contained twice as much half-cystine and a slightly higher proportion of basic amino acid than gamma-RSCP from RB.


T cell memory for the cytotoxic response to hapten-modified target cells. This study describes the development of memory and cytotoxic murine T cells against syngeneic haptne N equals[N-(3-nitro-4-hydroxy-5-iodophenyl-acetyl)-Beta-alanylglycylglycyl] associated antigen. Memory activity in this system had the following characteristics. a) In vitro challenged cells primed in vivo resulted in an augmented cytotoxic response compared to cells primed in vitro. b) The augmented cytotoxic response in vitro was antigen-specific for both target cells in the lytic reaction and stimulator cells in the secondary response. c) Memory activity was long lasting (at least 2 months). d) Memory cells were not cytotoxic. e) Memory activity as well as the cytotoxic cells generated in a secondary response in vitro were T cell dependent, These findings are consistent with the results of others who have investigated T cell dependent memory in other cell-mediated reactions.


Delayed hypersensitivity and granulomatous response after immunization with protein antigens associated with a mycobacterial glycolipid and oil droplets. A myocardial glycolipid (P3) mixed with protein antigens in oil-in-water emulsion induced lasting delayed hypersensitivity (DH) and granulomatous inflammation after intradermal injection into guinea pigs. This did not occur when P3 and bovine serum albumin (BSA) were given in Freund's incomplete adjuvant. The oil-in-water emulsions consisted of microscopic oil droplets suspended in aqueous medium. By separating oil and aqueous phases from BSA + P3 emulsion it was shown that antigen retained with oil droplets led to DH and granuloma formation. The association of antigen with oil droplets was P3 dependent and was quantitated with 125I-labeled BSA. The same phenomenon occurred with 125I-labeled rabbit gamma-globulin (RGG) + P3 emulsion. Fluorescein-conjugated RGG was observed in a particulate state within or on oil droplets in emulsion containing P3. These physical characteristics of antigen + P3 emulsion appeared to be important for immunogenicity.


Properties of antigen-specific suppressive T cell factor in the regulation of antibody response of the mouse. II. In vitro activity and evidence for the I region gene product. An antigen-specific suppressive T cell factor, which was extracted from carrier-primed T cells, was further characterized in an in vitro secondary antibody response. The factor was capable of suppressing secondary IgG antibody response of primed spleen cells when it was added to the culture together with relevant antigen. The suppressive T cell factor was not released from primed T cells by a short-term culture with antigen, but was kept bound to the membrane of the residual cultured cells, only the physical disruption of which can release the T cell factor. The target of the suppressive T cell factor was determined as being the helper T cell, since the factor did not exert any effect in the absence of the helper T cell with identical specificity to that of the factor. The suppressive activity was completely absorbed with alloantisera specific for products of the I region of H-2 complex, although various anti-immunoglobulin antisera failed to do so. Close analysis of the specificity of alloantisera capable of absorbing the suppressor molecule indicated that the suppressive T cell factor may, in fact, be an I region gene product probably coded for by genes in I-A and/or I-B (including I-E) subregions.


Evidence for more than one Ia antigenic specificity on molecules determined by the I-A subregion of the mouse major histocompatibility complex. Ia antigenic specificities determined by the I-A subregion of the mouse major histocompatibility complex have been examined in strain B10.D2 (H-2d), C57BL/10 (H-2b), and in a (C57BL/6xDBA/2) hybrid (BDF1; H-2b/d). Detergent solubilized, 3H-leucine-labeled antigen preparations were mixed with appropriate alloantisera and precipitation was induced either by addition of goat anti-mouse gamma-globulin or by addition of protein A-bearing Staphylococci. Sequential precipitation analysis showed that in strain B10.D2, Ia specificities 8 and 11 were co-precipitable, and that in strain C57BL/10, Ia specificities 8 and 9 were co-precipitable. In contrast, precipitation of specificities 9 and 11 from a BDF1 antigen preparation showed that these two Ia specificities were on separate molecules. The genetic implications of these data are discussed.


Isolation of stable and unstable ("suppressed") Beta2-microglobulin-deficient clones from a lymphoid cell line. After selection with anti-Beta2m and C against a human lymphoid line, about 10% of surviving clones manifested decreased cytotoxic sensitivity to anti-Beta2m("suppression") which gradually reverted to normal after 35 to 55 cell doublings. Antiserum alone induced suppression. Exposure of suppressed clones to DMSO substantially reversed suppression. One clone, surviving selection, had a stable decrease in anti-Beta2m sensitivity; it possibly represents a Beta2m mutation.


Analysis of cell-surface markers with staphylococcal protein A: fluorescence studies on mammalian lymphoid determinants. Membrane antigens including different classes of immunoglobulins, transplantation antigens, beta2-microglobulin, T lymphocyte specific antigens, and virally determined surface components were investigated using fluorescein-labeled Staphylococcal protein A in combination with cytofluorometric studies. Lymphocytes of seven species: mouse, rat, guinea pig, pig, cow, monkey, and human, and of ten human lymphoma-derived lines were tested. Analysis of the differential expression of surface markers revealed a reproducible reaction of protein A with cell-surface Fc of IgG actively produced by lymphoid cells from human, monkey, guinea pig, and pig, and with passively attached IgG molecules in the form of antibodies, directed against cell surface antigens of all lymphoid cells tested. No surface Ig was detected on so-called T lymphocytes. The distribution of cell-bound Ig density among surface Ig-positive cells was found to be different depending upon the origin of the cells with regard to lymphoid organ; it was parallel among the lymphoma lines tested and on peripheral blood cells from human, monkey, and pig, although large variations in fluorescence intensity among individual cells and among the different lines were recorded. Beta2-microglobulin determinants were found equally well on enriched human T and B cells. Transplantation, and T lymphocyte-specific antigens were detected on the majority of the lymphoid cells and on a restricted population respectively.


The role of radiotherapy in the treatment of primary tumours of the frontal sinuses. Three cases of primary frontal sinus carcinoma are described, all three in males. Radiotherapy for these tumours can be applied either combined with surgical treatment pre, or postoperatively using supervoltage radiation, or by itself. If applied alone, it may either be a palliative measure in inoperable cases, or a curative measure in radiosensitive tumours.


Immunochemical analysis of water-soluble antigens of chick retina in the course of embryogenesis. Water-soluble antigens of chick retina were investigated using rabbit antisera to total extract and to individual electrophoretic fractions of retinal extract by methods of immunoelectrophoresis and Ouchterlony precipitation test. In the retina of the adult chick six serum and eleven tissue antigens were demonstrated. The tissue antigens of the retina comprised one organ-specific antigen and ten inter-organ antigens which were characterized by nonuniform distribution in tissues and organs of adult chick. Three antigens out of these were found only in tissues of the eye (retina, iris) and in the brain--inter-organ antigens of 'narrow' specificity. The ohter seven inter-organ antigens were found in tissues of brain and eye, as well as in various tissues and organs of hens--inter-organ antigens of 'broad' specificity. A high degree of antigen similarity between retina and iris was observed. Anti-retina sera in chick lens could detect only inter-organ antigens of 'broad' specificity. In the course of embryogenesis the first to appear in the developing retina were inter-organ antigens of 'broad' specificity (on 3rd day of incubation). Formation of antigens of this group was completed by the 9th day of incubation. On the contrary, inter-organ antigens of 'narrow' specificity appeared later, in the period of histogenesis of retina (from 5 to 18 days of incubation). The organ-specific antigen of retina was found by 7th day of incubation. One of the inter-organ retinal antigens of'narrow' specificity (retina-iris-brain) appeared in the developing chick brain at the same time as in retina--on 10th-11th day of incubation. Using the indirect immunofluorescence antibody technique this antigen was identified in the cytoplasm of retinal cells and brain neurones, but was not detected in the nerve fibres.


Immunochemical characterization of the "native" type III polysaccharide of group B Streptococcus. The type III polysaccharide of -roup B Streptococcus has been isolated and purified by a method that employs washing of intact cells at neutral pH. That the polysaccharide prepared by this procedure is the "native" type III antigen is suggested by its molecular size in excess of 10(6) daltons, its degradation by acid and heat treatment to a fragment with immunologic characteristics of the classical HCl antigen, and its type-specific serologic activity. The type III polysaccharide in native form contains sialic acid, galactose, glucose, glucosamine, heptose, and mannose. It is acidic in nature, is resistant to neuramindiase degradation, contains no O-acetyl groups, and does not share antigenic determinants with capsular type K1 antigen of Escherichia coli or Group B polysaccharide antigen of Neiserria meningitidis.


Detection of alloantigens during preimplantation development and early trophoblast differentiation in the mouse by immunoperoxidase labeling. An immunoperoxidase-labeling technique allowing visualization of antibody binding to the cell surface at the electron microscopical level has been employed an an analysis of H-2 and non-H-2 alloantigen expression on the early mouse embryo. The presence of non-H-2 antigenic determinants has been confirmed on eight-cell, morula, and blastocyst stages of development. Contrary to previous reports, however, low levels of H-2 antigen have also been detected on the blastocyst. This is the earliest stage at which H-2 has been shown to be expressed on the fertilized mouse egg and may reflect the greater resolution of the immunoperoxidase technique. Using two different models to study the critical peri-implantation stages, those of experimentally induced blastocyst activation and blastocyst outgrowth in vitro, it has been demonstrated that antigen loss occurs on the trophectoderm at the time of implantation, and that this is not necessarily dependent upon maternal influence. It is suggested that the loss may be an important factor in the prevention of maternal immune rejection during the establishment of the fetal allograft. The two major components of the early postimplantation conceptus display a striking differential in antigenic status. The embryonic sac shows a high degree of peroxidase labeling, while the ectoplacental cone trophoblast is unlabeled. These findings add support to the concept of antigenic neutrality of the early trophoblast and its role in the maintenance of a normal fetomaternal immunological equilibrium.


Immunochemical studies on blood groups LXII. Fractionation of hog and human A, H, and AH blood group active substance on insoluble immunoadsorbents of Dolichos and Lotus lectins. The purified lectins from Lotus tetragonolobus and Dolichos biflorus were coupled to Sepharose 2B to make insoluble adsorbents for purification and fractionation of blood group A and H active glycoproteins. With both adsorbents, hog gastric mucin A + H blood substance (HGM), purified by phenol-ethanol precipitation, yielded fractions showing only A, only H, or AH activities. The AH fraction was obtained when the adsorbent column was overloaded with HGM and its A and H specificities seem to be carried on the same molecules since they were not separable by chromatography on either column. However A and H specificities of blood group substance from the stomach of a presumably heterozygous individual hog were both on the same molecules as they too could not be fractionated on either column. Analytical properties of the isolated fractions were generally similar to those of the unfractionated material, the purfied A substances had a higher galactosamine/fucose ratio than did the H substances. Although the original A + H showed very little specific optical rotation, the separated A and H substances rotated positively and negatively, respectively. The lectin-Sepharose adsorbents have also proven useful in isolating A or H substances directly from the crude commercial hog gastric mucin. Blood group A2 substance from a human ovarian cyst yielded two fractions on the Lotus-Sepharose column; the effluent did not interact with the Lotus lectin but precipitated the Ulex and Dolichos lectins and anti-A, and appears to contain type 1 H determinants. The other fraction reacted with Lotus and Ulex lectin as well as with Dolichos and anti-A.


Specific adsorption of IgM antibody onto H-2-activated mouse T lymphocytes. Evidence is presented to support the contention that IgM demonstrable by surface immunofluorescent staining on H-2-activated T cells represents specifically adsorbed B-cell-derived alloantibody. T cells activated to H-2 determinants expressed surface IgM only when the progenitor cell populations contained B lymphocytes. IgM was not detected on T cells activated to determinants which fail to stimulate alloantibody production (e.g., M-locus determinants). In addition, IgM-negative H-2 activated T cells (derived from B-cell-depleted lymphoid cells), unlike M-locus-activated T cells, adsorbed IgM in a specific manner when incubated in vitro with "early bleed" antisera raised against the activating H-2 determinants.


A single genetic element in H-2K affects mouse T-cell antiviral function in poxvirus infection. Cell transfer experiments using mice with recombinant H-2 haplotypes were used to map the H-2 regions which must be shared by ectromelia-immune T-cell donors and virus-infected recipients for transfer of virus clearance mechanisms in the spleen. K- or D-region genes were necessary and sufficient; I-region genes  were not involved. The remainder of the mouse genome could be varied widely without impairing the efficacy of T-cell antiviral function, provided either a K or a D region was shared in the donor-receipient combination. A mutation in a single genetic element of the K region of the H-2 complex abolished the antiviral effect of immune T-cell transfer in a donor-recipient combination which shared the K end.


Modification of murine T-cell cytotoxicity by preimmunization with M locus and H-2 incompatibilities. The effect of preimmunization of CBA/H mice with M locus and H-2 incompatible lymphocytes was examined on the in vitro generation of cytotoxic cells to both types of targets. Preimmunization with M locus did not help to generate cytotoxic cells to M-locus targets; however, the anti-H-2 cytotoxicity of M-locus-preimmunized lymphocytes was suppressed between the 6th and 19th day after preimmunization. In contrast, preimmunization with H-2 incompatibility produced an amplified lytic anti-H-2 response. The possible mechanisms are discussed.


Synthesis and secretion of alpha2-macroglobulin by cultured human fibroblasts. The following observations indicate that cultured human WI-38 fibroblasts synthesize and secrete alpha2-macroglobulin into serum-free medium: (a) after incubation of cultures with [35S]L-methionine, a labeled protein appeared in the medium which was precipitated by antiserum directed against alpha2-macroglobulin; (b) after incubation of cultures with [35S]L-methionine, a major band of radioactivity detected by polyacrylamide gel electrophoresis of the proteins in medium co-migrated with alpha2-macroglobulin; and (c) the amount of alpha2-macroblobulin in the medium, estimated both functionally and immunologically, increased with time in normal but not not puromycin-treated cultures.


Modulation of regulatory mechanisms operative in the cyclical production of antibody. Modulation of the cyclical response in rabbits to aggregated human gamma globulin (AHuIgG) was investigated in order to study some of the parameters involved in self-regulation of the immune response. Several mitogens (lipopolysaccharide [LPS], phytohemagglutinin [PHA], and concanavalin A [Con A]), when injected simultaneously with antigen, have been shown to modulate the normal splenic plaque-forming cell (PFC) response in rabbits to a single intravenous injection of AHuIgG. This response to AHuIgG has previously been characterized by the initial appearance of PFC in the spleen 3 days later, with a peak of PFC at 5 days after injection. The number of PFC in the spleen then decreases and remains at a low level until a second increase begins on day 10, peaking on day 13. The 8-day cycle between peak PFC repeats, with a third peak appearing on day 21. In the present studies, injection of LPS with AHuIgG was shown to affect the PFC response by enhancing only the initial peak of PFC, PHA was shown to enhance both the initial and secondary peaks of PFC, while injection of Con A with AHuIgG resulted in a prolonged increase in PFC with no apparent cycling. Irradiation 24 h after injection of antigen resulted in PFC kinetics similar to those observed with PHA, although the increase in PFC was more marked with irradiation. Thus, although LPS, PHA, Con A, and irradiation markedly affected the immune response to AHuIgG, Con A was the only substance which altered the cyclical appearance of PFC to HuIgG. The cyclical nature of the PFC kinetics was shown to occur with either intravenous or intraperitoneal injection of antigen and in both primary and secondary responses, provided that the rabbits were primed with a low dose of antigen. Data were obtained that suggest that the response in distal lymph nodes may be regulated by immunological events occurring in the spleen. Cycling of PFC was not observed in the draining node after subcutaneous injection of AHuIgG in the hind foot. However, if the antigen was also injected intravenously at the same time as the subcutaneous injection, the response in the node became cyclical.


Antigen-induced aggregation and modulation of receptors on hapten-specific B lymphocytes. Mouse spleen cells were subjected to a fractionation procedure designed to enrich for 4-hydroxy-3-iodo-5-nitro-phenylacetyl (NIP)- or DNP-specific B lymphocytes, which depended on adherence of specific cells to a layer of hapten-gelatin at 4 degrees C, recovery of bound cells by melting, and digestion of adherent antigen by collagenase. A population of cells resulted which contained 90% typical B cells and 37% of cells capable of binding a fluorescent, haptenated polymeric protein. Fractionated cells were reacted in vitro with fluorescent conjugates of the specific haptens with polymerized flagellin [NIP-polymerized flagellin (POL)-tetramethylrhodamine isothiocyanate conjugate or DNP-POL-fluorescein isothiocyanate conjugate] under a variety of conditions, with the aim of investigating the behavior of Ig receptors on B lymphocytes after exposure to antigen; Experiments were performed with immunogenic and tolerogenic concentrations of antigen. Furthermore, four experimental designs were used, namely: (a) brief labeling with fluorescent antigen followed by culture without antigen (pulse design); (b) culture in the continuous presence of fluorescent antigen (continuous-labeling design); (c) culture in the continuous presence of nonlabeled antigen followed by labeling of unoccupied receptors by fluorescent antigen (receptor status design); and (d) culture with nonlabeled antigen for 2 h followed by incubation without further antigen for 20 h and labeling with fluorescent antigen (modulation design). Further insight into receptor occupancy and distribution was gained by the use of fluorescent antihapten and antiglobulin reagents. It was found that both immunogenic and tolerogenic antigen concentrations caused rapid patching and capping of the receptors to which they attached, followed by endocytosis and probably some shedding of Ig receptors. However, a proportion of cells continued to bear some cell surface antigen for 24 h. The immunogenic antigen concentration failed to completely remove the receptor coat from the cell surface. At all stages of immunogenesis, plentiful unoccupied receptors could be demonstrated. The tolerogenic concentration nearly saturated available receptors, and in its continuous presence, only few unoccupied or antigen-occupied surface receptors could be detected after 24 h of culture. Experiments of the modulation design showed that brief incubation with the tolerogenic concentration appeared to suppress receptor resynthesis, as few new receptors could be demonstrated after 20 h of further culture without antigen. Experiments were performed to determine whether fractionated cells prepared from spleens of 8-day-old mice showed an unusual tendency for modulation, even with immunogenic antigen concentrations. They were found to behave essentially like adult fractionated cells. The results are discussed in the framework of current theories of B-lymphocyte activation and tolerization.


Structural characteristics of the alloantigens determined by the major histocompatibility complex of the guinea pig. The GPLA B and Ia (I region-associated) antigens are the products of genes found in the guinea pig major histocompatibility complex. Because of their importance in immune response phenomena, a structural study of these antigens was undertaken. [3H]leucine and [3H]fucose were internally incorporated into guinea pig lymph node cells. The GPLA B and Ia antigens were solubilized by the nonionic detergent Nonidet P-40, purified by affinity chromatography using an adsorbent column of lentil lectin, isolated by immunoprecipitation, and examined by discontinuous polyacrylamide-sodium dodecyl sulfate gel electrophoresis. The GPLA antigens B.1, B.2, B.3, and B.4, were shown to be glycoproteins of mol wt 40,000 daltons and to be noncovalently associated with a 12,000 dalton protein. The molecules bearing B.2 and B.3 in a B.2/B.3 heterozygote are shown to be separable, suggesting the antigenic determinant is a primary gene product. In addition, a new GPLA determinant, S, which resembles the B antigen in that it is found on a molecule of approximately 40,000 daltons, was studied. In a B.2/B.3 S+ animal the molecule bearing antigen S was shown to be independent of those bearing B.2 and B.3, providing evidence that the genes determining B and S are at separate loci. The Ia-bearing molecules identified by anti-Ia.2,4 are glycoproteins of mol wt 58,000 daltons which are composed of two subunits of 33,000 and 25,000 daltons, respectively, linked by disulfide bonds. The Ia-bearing molecules are independent of GPLA-bearing molecules, indicating different loci determining these antigens. By all criteria, the guinea pig GPLA B antigens appear homologous to the murine H-2D and H-2K antigens, while the guinea pig Ia antigens appear homologous to the Ia antigens of the mouse.


Effect of recent antigen priming on adoptive immune responses. III. Antigen-induced selective recruitment of subsets of recirculating lymphocytes reactive to H-2 determinants. Information was sought on the reactivity of thoracic duct lymphocytes (TDL) from parental strain mice injected intravenously with large numbers of irradiated semiallogeneic spleen cells. TDL collected at 1 day after spleen cell injection were almost totally depleted of lymphocytes able to produce cell-mediated lympholysis (CML), a graft-versus-host (GVH) reaction, and skin allograft rejection against the H-2 determinants on the injected spleen cells. Normal or near normal responses were observed against third-party determinants. In the case of CML, there was no evidence that the unresponsiveness was due to suppressor cells. In marked contrast, the capacity of TDL to exert a specific mixed lymphocyte reaction (MLR) against the injected determinants was reduced by no more than two to fourfold; this applied whether MLR were measured in vivo or in vitro. Injection of normal rather than irradiated semiallogeneic spleen cells gave similar results. Complete and specific removal of MLR-producing lymphocytes was achieved, however, in a different system in which parental strain T cells were filtered from blood to lymph through irradiated F1 hybrid mice. Since this system presumably provided a much higher concentration of H-2 determinants to the responding lymphocytes, it is suggested that the differing results obtained with these two systems may indicate that certain cells reactive to H-2 determinants are of low affinity, their reactivity being detected in the MLR, but not by other parameters. With both systems, MLR-producing lymphocytes reappeared in the lymph after 2-3 days; the cells collected at this stage gave an MLR of altered kinetics. The present data, in toto, suggest that under certain conditions of antigen presentation, virtually all recirculating lymphocytes reactive to a given set of H-2 determinants can be induced to leave the circulation for a period of 1-2 days. After responding to the injected determinants (presumably in organs such as the spleen), the cells re-enter the circulation in an activated state after 2-3 days.


Specific binding of alloantigens to T cells activated in the mixed lymphocyte reaction. Immunoglobulin (Ig) is present on a large fraction of T cells from unfractionated lymphocytes activated by in vitro stimulation with H-2-incompatible cells (mixed lymphocyte reaction [MLR]). Removal of bursa equivalent-derived (B) cells from the responder cell population before mixed culture, by filtration through nylon wool columns, reduces the percentage of Ig-bearing responder T blasts to background levels. Thus, Ig on the T blast is probably of B cell origin. A large fraction of T blasts activated against the stimulator cells. This staining occurs with "early" and hyperimmune alloantisera, including the 7S fraction of the latter. B-depleted responder cells were activated against a mixture of two different stimulator cells and the resulting T blasts stained with different concentrations of sera directed either against one or both stimulator cells. We obtained results which strongly suggest that most or all responder T blasts stain with only one antistimulator serum. When antisera directed against different segments of the H-2 complex of the stimulator cells were used, it seemed that most responder T cells only bound antibody directed against a single segment. We propose that T cells activated in MLR carry stimulator alloantigens on their surface, and that this is due to specific antigen binding, not requiring the presence of B-cell-derived antibody. These histocompatibility antigen-binding T blasts can be detected by appropriate antistimulator alloantibodies.


Sheep red blood cell-specific helper activity in rat thoracic duct lymphocyte populations positively selected for reactivity to specific strong histocompatibility alloantigens. These studies show that positively selected T-cell populations, having enriched reactivity in the mixed lymphocyte interaction and the graft-versus-host reaction to strong alloantigens of a chosen major histocompatibility complex haplotype, also possess helper activity which is quantitatively normal in the generation of primary antibody responses to sheep red blood cells in vitro. Such positively selected populations give a linear dose plaque-forming cells response curve indistinguishable from that seen with normal unselected T-cell populations. These findings imply that T cells reactive to histocompatibility antigens also react to conventional antigens, and the possibility is raised that they may do so by some recognition mechanism involving multiple specificities.


Isoelectric focusing and electrophoresis of cerebrospinal fluid proteins in muscular dystrophies and spinal muscular atrophies. In the very few previous investigations of the CSF-proteins in muscular dystrophies the results have generally been reported as normal. In spinal muscular atrophies a barrier-damage pattern of CSF-proteins has been found in amyotrophic lateral sclerosis (ALS). In the present investigation the CSF-proteins were examined by isoelectric focusing and quantitative paper electrophoresis in 13 patients with muscular dystrophies and in 11 patients with spinal muscular atrophies. On isoelectric focusing, CSF-protein abnormalities were found in 85% of the cases with muscular dystrophies and in all patients with spinal muscular atrophies. Differences in the CSF-protein patterns were observed within the group of muscular dystrophies and between these and the cases of  spinal muscular atrophies. In ALS and in myotonic dystrophy, abnormal CSF-protein fractions occurred mainly in the alkaline pH-range, while in limb-girdle dystrophy and the patient with facioscapulohumeral dystrophy, aberrant fractions appeared mainly in the acidic region. CSF-protein abnormalities were found in both the alkaline fractions (HAFs) with pI 9.2-9.6 and a fraction with PI 7.1 were found in half of the patients with myotonic dystrophy. The CSF electrophoresis in myotonic dystrophy showed increased levels of beta1-globulin in all cases examined. Signs of barrier-damage were commonly encountered in ALS in contrast to the muscular dystrophies, except for myotonic dystrophy. The results are discussed in terms of possible diagnostic value and with regard to pathogenetic significance, particularly in relation to the current hypothesis of a neural involvement in muscular disorders.


Clinical observations of juvenile nonprogressive muscular atrophy localized in hand and forearm. Twenty-seven patients with juvenile nonprogressive muscular atrophy localized in the hand and forearm were analyzed. The clinical characteristics were juvenile male occurrence, insidious onset, specific distribution of localized muscular atrophy and a stationary course. On electromyography, denervation voltage (or giant NMU) is found in the atrophied muscles and sometimes in contralateral nonatrophied ones. Sensory disturbance was not remarkable. Although the etiological factor was not known, strenuous exercise of arms in sports was noted frequently in the history.


Dynamic aspects of peripheral nerve changes in progressive neural muscular atrophy: light- and electronmicroscopic studies of serial nerve biopsies. Serial nerve biopsies were performed at an early, and at an advanced stage of the disease in 2 patients with progressive neural muscular atrophy. The early biopsy showed a complete loss of the large diameter and thickly myelinated fibres, as well as an expansion of the endoneurial interstitium in both cases. Myelinated and unmyelinated fibres exhibited axonal degeneration in all biopsies occasionally. "Onion bulb" formation, a typical feature of peripheral neuropathy in neural muscular atrophy, was found to be prominent only in the latter biopsies. As regards the formal pathogenesis of hypertrophic neuropathy in neural muscular atrophy, axonal dystrophy and interstitial changes of the endoneurium were regarded as primary phenomena, demyelination and "onion bulb" formation as secondary. A possible causal relation between axonal dystrophy and interstitial changes, observed in these cases, is discussed in the light of the present literature.


Histochemistry and electron microscopy of muscle fibres in a case of congenital paramyotonia. In a case of congenital paramyotonia a muscle biopsy was performed and studied morphologically, histochemically and ultrastructurally. A clearcut pattern of changes has been observed with ATPase and oxidative enzymes. On electron microscopy special changes known as "tubular aggregates" were found. The relationship between the two findings, as well as the significance of such alterations in the range of periodic paralyses and myotonic phenomena, are discussed.


Biochemical findings in multiple sclerosis. II. A detailed study of the serum IgA, IgG and IgM levels of 772 MS patients compared with 226 neurological controls. In the second paper of this series, a detailed study of the three main serum immunoglobulins for a group of 772 MS patients is compared with a group of 226 neurological controls. The results are studied according to sex and different age groups. Except for a slight elevation of the IGM levels in the MS group, especially between 40 and 60 years, no clear cut distinction between the two groups of patients was found.


[Intracranial and spinal hemorrhage in haemophilia (author's transl)]. Among 234 hemophiliacs, 14 patients (5.9%) with intracranial and 1 patient (0.4%) with intramedullary hemorrhage were observed and examined. The average age at the time of hemorrhage was 17 years in patients with severe hemophilia and 43 years in patients with moderate hemophilia. The patients with mild hemophilia and intracranial bleeding were 5 and 58 years old, respectively. Eight intracerebral, 3 subdural and 4 subarachnoid hemorrhages were encountered. In 7 cases bleeding was posttraumatic and in 5 it occurred spontaneously. In 2 patients hemorrhage followed a sudden increase of intracranial pressure. In 1 patient reliable data could not be obtained. Four of the 8 patients with intracerebral bleeding died; all survivors displayed some residual neurological abnormalities. All patients suffering from subarachnoid bleeding recovered completely. No serious complications were encountered after 3 lumbar punctures, 5 cerebral angiographies and after 1 myelography. Only in 1 case with an acquired inhibitor did a local hematoma develop at the site of puncture for the angiography. The same diagnostic procedures should be performed after substitution therapy with anti-hemophiliac plasma in a hemophiliac without inhibitor as in non-hemophiliacs if intracranial or intramedullar bleeding is suspected. In hemophiliacs with an anticoagulant, angiography or lumbar puncture should be performed only in exceptional cases.


Evaluation of results of thymectomy in myasthenia gravis. The results of thymectomy carried out in 150 cases of myasthenia gravis are discussed. In a group of 123 cases followed for 1 to 5 years after the operation, full remission was observed in 24.4% of cases, significant improvement in 36.6%, slight improvement in 24.4% and no improvement in 8.1%, while deterioration occurred in 1.6% of patients. No correlation was found between the result of the operation and the age and sex of patients, but better results were achieved in those treated surgically rather soon after the onset of symptoms. This correlation was particularly evident in the group with full remissions. The results obtained in the cases without thymic tumors were better than in the cases with tumors. No correlation was noted between the results of the operation and the histological characteristics of the thymus in the group with thymic hyperplasia and in the group with thymic atrophy. The surgically treated group (150 cases), compared with the conservatively treated group (75 cases), showed the superiority of the surgical method (lower rate of death and deterioration, higher rate of improvement and remission). In discussing the indications for surgical treatment the authors emphasize that advances in anaesthesiology in recent years have reduced the risk of operation. It is suggested that the indications for surgical treatment should be expanded and operations should be performed as early as possible after the onset of clinical manifestations without regard to the age and sex of the patient. Operation should not be considered in cases belonging to group 1, 2a (sometimes 2b) only, with duration of the disease over 8-10 years and with little or no progression of the process, if the presence of a thymic tumor has been excluded.


Investigations on the mechanism of nocturnal epilepsy. Sixty polygraphic all-night sleep tracings were obtained, including 40 from 20 patients with epileptic attacks occurring during sleep (during treatment with anticonvulsants and after their withdrawal), and 10 tracings in each of two groups serving as controls: a group of patients with seizures occurring in the waking state and a group of healthy controls. A significant difference was observed in the sleep pattern of the patients with nocturnal attacks (who were good sleepers and received no anticonvulsants) and healthy controls. These patients had a significantly higher proportion of III + IV stages of the slow-wave sleep phase. Besides that, in all epileptics disruption and lability of the REM phase was observed, without a statistically significant reduction in the proportion of its duration during the whole sleep. Clinical seizures developed usually in the SW phase but one seizure occurred during the REM phase. The effects of anticonvulsants on sleep EEG tracings require further investigation. A significant difference was observed in the morphology and dynamics of EEG changes in both groups of epileptics. The patients with nocturnal attacks had much more frequently normal tracings in the waking state. During sleep focal changes were often found. Seizure activity of the spike-wave type was less frequent than in patients with waking attacks and the dynamics of this activity were different. It seems possible that the pathways of spread of epileptic discharges are different in these two groups of epileptics.


[An intracranial epidermoid caused by trauma (author's transl)]. A report of an epidermoid caused by trauma. The traumatic genesis was conclusively demonstrated in the course of an almost uninterrupted 28-year observation. The differential diagnostic difficulties and the consequences following therefrom are emphasized.


Lanthanum staining of cell surface and junctional complexes in normal and malignant human oral mucosa. Biopsies from normal and malignant human oral epithelium were studied electron histochemically using lanthanum nitrate for a selective demonstration of junctional complexes at the plasma membrane of adjacent cells. The heavy metal complex was found to settle irregularly, sometimes in intimate proximity to the epithelial cell, in areas corresponding to the glycocalyx, gap junctions, desmosomes and nonspecific contact areas. Tight junctions were rarely seen. The lack of these morphological adhesive areas, the desosomal detachment seen between cancer cells and the increased lanthanum-positive cell surface may be possible factors responsible for the dissociation of oral malignant cells.


The immunohistochemical identification of hormone-producing cells in formalin-fixed, paraffin-embedded human pituitary tissue. A method is described, combining immunohistochemistry and histological staining methods in the same section, for the identification of ACTH- and STH-producing cells in formalin-fixed, routine processed human pituitary tissue. The results indicate that this method is suitable to employ in practice.


A quantitative morphological study of the carotid bodies of rats living at a simulated altitude of 4300 metres. A quantitative histological study was carried out on the carotid bodies of 10 normal rats and 10 rats living in a hypobaric chamber at a pressure of 460 mm Hg from 25 to 96 days. In the chronically hypoxic rats there was a four-fold increase in the mean combined volume of the carotid bodies. Morphometric analysis disclosed a three-fold increase in the mean volume of specialised glomic cells and a ten-fold increase in the mean volume of capillaries, although the proportion of glomic cells was actually significantly decreased. In all our hypoxic rats there was evidence of both right and left ventricular hypertrophy. However, there was no linear relation between total carotid body volume or volume of glomic cells on  one hand and the right and left ventricular weight, on the other hand. Although there was no linear relation between combined total carotid body volume and duration of hypoxia, the linear relation between glomic cell volume and duration of hypoxia was significant at the 5 per cent. level. The increase in vascularity of the hypoxic carotid body may be a mechanism to increase blood flow and thus oxygen transport to a hypoxic organ with increased metabolic activity. Small quantities of an amorphous hyaline material of unknown nature were found in relation to capillaries and type I cells in all the hypoxic rats.


Retention and redistribution of proteins in mammalian nerve fibres by axoplasmic transport. Fast axoplasmic transport is characterized by a crest of labelled activity moving down nerve fibres after injection of the L7 dorsal root ganglion with the amino acid precursor (3H) leucine, the crest followed by a plateau which represents in part a later egress of labelled components from compartments in the cell bodies and in part materials left behind the advancing crest. 2. after making ligations just below the ganglia at different times after injection of the precursor, a small downward slope of locally retained activity of incorporated materials is seen in the plateau remaining in the nerves. The slope becomes changed to a horizontal level when in addition a distal ligation is made as a result of the redistribution of labelled materials within the doubly ligated nerve segments. 3. the outlfow pattern at later times, at a day and longer after injection, shows an additional spread of activity from the cell body region. The pattern of outflow gradually levels off at later times as additions of activity are made to the more distal part of the nerves. The activity retained in the nerves becomes less free to become redistributed in the course of several days. 4. The temporal changes in the outflow patterns can be accounted for by the local retention and redistribution of the labelled materials within the fibres. Later additions of labelled materials compartmented in the cell bodies also contribute to the later pattern of outflow. A "unitary" view for fast and slow transport is presented based on the transport filament hypothesis earlier proposed to account for fast axoplasmic transport.


Prosthetic support for unilateral facial paralysis. Removable prostheses were used to determine that esthetics and speech could be improved for patients with permanent unilateral facial paralysis. Esthetics had to be compromised somewhat to obtain the maximum benefit for intelligible speech. This procedure can be beneficial in helping patients who are high surgical risks or for whom surgery, for various reasons, is unacceptable.


Application of "cusp writer" findings to practical and theoretical occlusal problems. Part I. The cusp writer is a teaching aid used to facilitate understanding of the correlation between cusp angulation and the various occlusal determinants. Cusp writings represent an on-edge view of mandibular movement occurring in translation and rotation. Cusp writings demonstrate that: (1) the influence of incisal guidance on cusp angulation is almost 60 per cent greater in molars and 100 per cent greater in pre-molars than the influence of the condylar guidance; (2) steepening or flattening by wear, grinding, or reconstructing the lingual contour of the upper canine or other incisal guide teeth by as little as 1 mm. can change the incisal guide angulation from 15 to 25 degrees depending upon the vertical overlap; (3) these changes can create interceptive or deflective occlusal contacts with trauma and pain on the balancing side in the molar and temporomandibular joint areas; and (4) the incisal guidance of denture teeth is inevitably altered when teeth are tilted, uprighted, or turned for esthetics or phonetics. Such changes alter the occlusal function, harmony, and stability of the dentures.


[Transhepatic embolization of oesophageal varices (author's transl)]. Embolization of the aeso-cardiotuberositary varices, by selective catheterization of the coronary vein of the stomach and the posterior gastrics, seems to be valuable palliative treatment in portal hypertension, owing to its relative innocuousness, inspite of a certain number of failures. It is carried out by transhepatic puncture of the portal system or catheterization of the umbilical vein.


3H-thymidine autoradiography of the CSF cells in cases of non-neoplastic disease. Human CSF cells in cases of non-neoplastic disease of the central nervous system (CNS) were examined in vitro by 3H-thymidine autoradiography. Immediately after withdrawl by lumbar or ventricular puncture, the CSF was incubated in a sedimentation chamber at 37 degrees C for 1 hr with an admixture of 3H-thymidine at a concentration of 1-2 muCi/ml CSF. In a few cases the CSF withdrawn was incubated in a glass tube in the same condition as in a sedimentation chamber, and the CSF cells were collected by centrifugation. The CSF cells collected were fixed in methanol and the microautoradiographic procedure was performed. Labeled CSF cells were found in 21 cases out of 22. The average labeling index of the total nucleated cells was 0.22% with the highest labeling of 0.74%. Almost all the labeled cells were thought to be medium to large sized lymphocytes and monocytoids. Peripheral blood was examined by a similar method and the results were compared with those of the CSF. It may be noteworthy that thre exist DNA synthesizing cells in the CSF even in a non-neoplastic state of the CNS, although the number is not large.


[Anomalies of the pattern of lumbosacral nerve roots and its clinical significance (author's transl)]. Twenty personal observations and 18 cases collected from the literature are analysed. The most frequently encountered anomaly were: common dural origin of 2 nerve roots and common exit of 2 nerve roots through the intervertebral foramen. Other anomalies comprised: interradicular connections and Y-shaped or horizontla course of the nerve root. Multiple anomalies were not encountered. In 9 out of 20 patients in the own series and in 6 out of 18 patients reported in the literature, history and clinical findings suggested prolapsed intervertebral disc, operation revealed only nerve root anomalies. Decompression produced improvement or complete relieve of previously existing signs and plain X-rays is not possible. The diagnosis is based on myelographic findings. The pathogenesis of the anomalies is discussed. It is suggested that they should not be considered as a causative factor of low back pain or sciatica.


[The macrophage electrophoretic mobility LAD test--a diagnostic method for multiple sclerosis (author's transl)]. With the MEM test (Field) one can establish a cellular immune reaction becarus the sensitized lymphocytes release the macrophage slowing factor (MSF) upon interaction with the appropriate antigen. A macrophage migration inhibition was detected in some neurological diseases with destruction of the parenchyma. The modification MEM-LAD (linoleic acid depression) test made further differentiation possible in the 146 neurological patients and normals. The reduction of macrophage mobility inhibition was 94.7 + 4.7% in multiple sclerosis (MS) cases as compared with that of normals of 55.1 + 3.7% and of other neurological diseases of 47.8 + 7.1%. There were no significant differences due to the course and duration of the disease or to immunosuppressive therapy. The pathogenically important results in relatives of MS patients with values between the MS and normal group (78.5 + 0.7%) in mothers suggested a familial (genetic) disposition. The same value was found in a monozygotic twin of an MS patient. The results in the children studied showed that besides the endogenic metabolic component the aetiopathogenically important exogenic factors can operate early in life. In correlation with the principle of the MEM-LAD test the suppressive action of linoleic acid can result in a further therapeutic concept.


Electromyographic findings in the so-called non-progressive myopathies. Electrophysiological findings in 40 cases of non-progressive myopathies are reported, and compared with a group of 20 cases of Duchenne progressive muscular dystrophy and a control group. In all cases the electrophysiological changes were of the mild s. c. myogenic type. The involvement of proximal and distal muscles was equal without prevalence in proximal muscles as is typical for Duchenne's dystrophy. EMG reexaminations showed a slight progression of the diseases. A peculiar feature of myotubular myopathy was spontaneous activity (fibrillation) in 70% of muscles. A myogenic character of the process of congenital deficiency of muscle innervation with preserved number of motor units is suggested.


Neuropsychiatric symptomatology with chronic renal insufficiency in the stage of compensated and decompensated retention. I. CNS disturbances. 80 strictly selected patients with chronic renal insufficiency with plasma creatinine values of 1.4--14.5 mg% were examined according to a fixed scheme to determine the presence of symptoms and signs of renal encephalopathy. The general cerebral symptoms complained of were headache in 33.4% of the patient material, dizziness in 30.3%, easy fatigability in 62.5%, giddiness in 18.8% and insomnia in 37.5%. The most prominent neurological findings were hyperactive deep reflexes in 30% and action tremor in 23.8%. The symptoms of organic brain syndrome were impairment of memory in 32.5%, weakness of concentration in 28.8% and lability of affect in 63.7%. Diffuse EEG abnormalities were found in 26.2%. While the clinical neuropsychiatric symptoms did not show any statistically significant correlation with the various internal medical data, a trend was observed in the greater number of pathological EEGs with an increase in the impairment of renal function. Furthermore, there was a statistically significant correlation, (alpha less than or equal to0.015) between the occurrence of pathological EEGs and the plasma creatinine and BUN values. It is remarkable that the patients with abnormal EEGs had a relatively low mean creatinine level of 5.89 mg%. The strict dietetic management of the patients is regarded as one of the deciding factors for the relatively low frequency of neuropsychiatric symptoms in the material studied.


Neuropsychiatric symptomatology with chronic renal insufficiency in the stage of compensated and decompensated retention. II. Peripheral nerve disturbances. 80 strictly selected patients with chronic renal insufficiency with plasma creatinine values of 1.4-14.5 mg% were examined for clinical and electrophysiological signs of nephrogenic polyneuropathy. The motor symptoms complained of were cramps in 43.8% of the patients, "restless leggs" in 18.7%, muscular twitchings in 12.5%. It was emphasized that the first two symptoms do not always indicate the presence of polyneuropathy. 30% complained of paresthesias, 5% of "burning feet". The most frequent clinical finding was the impairment of vibration sense in the feet in 37.5% followed by diminshed appreciation of passive movement of the toes in 30%, weakening or absence of the ankle jerk in 23.8% and finally, weakening of the patellar reflex in 5%...


[The EEG after frontal and frontobasal skull and brain injury (author's transl)]. Eighty-eight patients with confirmed frontal and frontobasal lesions were examined. The EEG changes were investigated according to type and frequency both after recent trauma and in the later stages (Fig. 1). The general changes showed mainly slight (Fig. 2B) and only rarely severe, dysrhythmias (Fig. 3A) which, however, were frequently associated with paroxysmal outbursts. Focal findings were largely temporal and frontal in the second place (Fig. 2A). All pathological EEG changes showed a tendency to improvement, so that in the late phase they occurred rarely and less severe (Fig. 4). They showed no essential difference from the EEG of uncomplicated craniocerebral traumata. The relations between the EEG and the clinical form of craniocerebral trauma were examined. In contusions of the head the normal EEG was predominant. The pathological EEG was prominent with cerebral concussion and even more so with cerebral contusion. On the other hand, there was no relationship between the EEG and the type of fracture. A significant predominance of pathological EEG was found only in frontobasal fractures with rhinorrhoea. The EEG changes, classified as focal and dysrhythmic, show no statistically significant relationship either to the form of craniocerebral trauma or to the type of fracture. The effect of force coming form in front has apparently been used up chiefly in producing the frontal or frontobasal fractures, and leads to relatively slight cerebral damage. The clinical significance of the EEG does not actually lie in the diagnosis of these injuries, but, as always, in the establishment of the severity and extent of the cerebral lesion and the detection of complications.


Carcinoma of the esophagus. Long-term results. Between 1940 and 1967, we treated 609 patients with primary carcinoma of the esophagus by three different methods. In the first group of 170 patients, surgical resection predominated. In the next 166 patients, radiation therapy was used as the primary method. In a third group (135 patients) and a fourth group (138 patients), radiation therapy was followed by resection in the operable cases. After comparing the results of the three methods, we believe that radiation therapy followed by resection produces the best results. Moreover, this approach is justified by the discovery of residual carcinoma in most surgical specimens from patients who have had radiation therapy.


Large-dose bleomycin therapy and pulmonary toxicity. A possible role of prior radiotherapy. Bleomycin sulfate pulmonary toxicity was encountered in nine of 101 patients receiving high-dose therapy for widespread testicular cancer. The pulmonary presentation was separable into two categories: (1) an early or minimal from with dyspnea on exertion, minimal roentgenographic findings, and normal arterial partial pressure of oxygen at rest and (2) a severe form, with prominent roentgenographic findings and hypoxemia at rest. All five patients with the severe form died, while the remaining four patients with the minimal presentation recovered. Prior thoracic radiotherapy appeared to predispose to bleomycin pulmonary toxicity, as this complication developed in five of 12 patients receiving prior chest radiotherapy vs four of 89 not receiving radiotherapy (p less than .001). The fatality rate of 5% with high-dose bleomycin therapy is acceptable in view of the 75% response rate and substantially improved survival achieved with bleomycin combination chemotherapy in metastatic testicular cancer.


Epiloia in twins: a problem in diagnosis and counselling. A family is reported in which the sixth and seventh children, who are twins, have epiloia. No other member of the family has any of the accepted stigma of this condition. It is shown that the twins are dizygotic and so are likely to have inherited their epiloia. Biochemical studies suggest that the father has epiloia and, in view of this, pigmented skin lesions assume diagnosis importance. The significance of this is discussed both in relation to diagnosis in general and to genetic counselling in this family where one of the other children might be affected.


Improved chemotherapy for small-cell undifferentiated lung cancer. Bleomycin sulfate, doxorubicin hydrochloride, cyclophosphamide, and vincristine sulfate combination chemotherapy was given to 29 patients with small-cell undifferentiated lung cancer. Only four of these patients had limited disease, and in these patients there was 100% complete remission; two of these four patients remain in complete remission at more than 52 and 60 weeks. Of the 25 patients with generalized disease, 18 (72%) had neoplasm regression (greater than 50%), including two with complete remission (8%). The median duration of remission was 25 weeks. The median survival time from diagnosis was 39 weeks and that from initiation of therapy, 35 weeks. The drug regimen was well tolerated, and although substantial leukopenia was produced, there were only three patients in whom granulocytopenic infections developed. There was only one drug-related death.


Fatal pulmonary reaction from low doses of bleomycin. An idiosyncratic tissue response. There were two cases of fatal interstitial pneumonia secondary to bleomycin sulfate administration. Although bleomycin pulmonary toxicity is generally thought to be dose-related and occurs infrequently with a total cummulative dose less than 300 to 400 units, the two reactions reported here occurred with doses of 105 and 165 units. Fatal bleomycin-induced pneumonia has been previously reported at these low dosages, and physicians should be aware that this toxic reaction may occur as an idiosyncratic response. Previous thoracic irradiation may be a predisposing factor. Patients receiving bleomycin should be meticulously monitored by interrogation for cough, dyspnea, and chest pain; by auscultation for rales; by serial chest roentgenograms; and by determinations of vital capacity and single-breath carbon monoxide diffusing capacity.


The endoscopic appearance of the duodenum in Whipple's disease. Duodenoscopic examination of a patient with Whipple's disease revealed that the duodenal mucosa appeared to be partially covered with a yellow-white material. On closer inspection, however, this was found to consist of enlarged villi interspersed with normal-looking mucosa. Histologically these enlarged white villi were shown to contain a heavy accumulation of lipid and typical PAS positive macrophages. The duodenal mucosa was endoscopically normal and histologically showed only a few PAS positive macrophages one year after instituting antibiotic therapy.


[Biogenic amines in endocrine cells producing polypeptide hormones. The APUD-cells (Pearse) (author's transl)]. APUD-cells (A = Amine, P = Pprecursor, U = Uptake, D = Decarboxylation) are endocrine cells with numerous common characteristics: 1. They produce polypeptide hormones of low molecular weight. 2. They possess specific 1-amino acid decarboxylases to synthesize catecholamines and/or 5-hydroxytryptamine from precursors. 3. A part of the APUD-cell family primarily accumulates biogenic amines within their cytoplasm. 4. All APUD-cells are rich in specific secretory granules that are the morphological substrate for most of their common features.--Enzyme histochemically the APUD-cells represent a rather heterogenous population. The capacity to take up biogenic amines from the blood is confined to a smaller numer of APUD-cells. The concept of the APUD-cells representing a peripheral neurosecretory system (Pearse und Polak, 1971) appears to be premature. It is probable that in several APUD-cells the accumulation of biogenic amines only constitutes a phenomenon without any physiological relevance which, notwithstanding, could explain pathological reactions of APUD-cells.


Cytochemical localization of catalase and peroxidase in sinusoidal cells of rat liver. The cytochemical localization of catalase and peroxidase in various sinusoidal cells of rat liver, i.e., in Kupffer cells, endothelial cells, and fat-storing cells has been investigated. The alkaline 3,3'-diaminobenzidine technique reveals distinct "catalase-positive particles" in all three cell types. The particles are round to oval-shaped, measuring 0.1 to 0.3 mum. in diameter. The diaminobenzidine reaction product is distributed uniformly over their matrix, often obscuring the distinct limiting membrane. In fat-storing cells the particles appear in close proximity of lipid droplets. No evidence of fusion of the limiting membrane of the particles with that of phagolysosomes containing latex particles was observed. The "catalase-positive particles" appeared often in close proximity of endoplasmic reticulum, but by examining consecutive serial sections we could not find any convincing evidence of direct continuity between the two organelles. In addition to catalase there is an endogenous peroxidase in the endoplasmic reticulum and nuclear envelope of Kupffer cells. Whereas peroxidase is sensitive to aldehyde fixation and has its optimal pH in the neutral range, the staining for catalase requires prior fixation with glutaraldehyde and isoptimal at pH 10.5. By using proper fixation and incubation conditions the two enzymes have been visualized selectively, and it is demonstrated that they occupy two distinct intracellular compartments within the Kupffer cells: the catalase in the matrix of particles and the peroxidase in the endoplasmic reticulum. The possible functional role of catalase in various sinusoidal cells is briefly discussed.


Observations on small granule cells in adult human bronchial epithelium and in carcinoid and oat cell tumors. Small granule cells are described in the segmental bronchi of adult humans. Rarely, these cells were seen abutting the lumen of the airway. A luminal location has been described for these cells in the bronchial epithelium of human fetuses and neonates and it is suggested that luminal small granule cells may be a rare, although normal component of the adult human bronchial epithelium. The dense core granules of the small granule cells stained intensely with phosphotungstic and at low pH suggesting that they contain glycoprotein. Granules present in a carcinoid and an oat cell tumor stained similarly. The role of the small granule cell in the genesis of bronchial carcinoid and oat cell tumors is recognized.


A morphologic and biochemical study of the fate of antibody-bearing liposomes. The effect of the intravenous administration of gamma-globulins, entrapped or not within liposomes, was investigated in rodent liver. Desialation of gamma-globulins did not influence significantly their uptake by this organ. Fluorescein-labeled nonspecific gamma-globulins and antibodies raised against the lysosomal acid alpha-glucosidase were entrapped within two types of liposomes. The administration of negatively charged liposomes made of lecithin, cholesterol, and dicetylphosphate was badly tolerated. Positively charged liposomes, in which dicetylphosphate was replaced by stearylamine, allowed a considerably higher yield of entrapment and were well tolerated by the animals. Approximately half of the fluorescent gamma-globulins were recovered in the liver, and evidence is presented for the intralysosomal localization of positively charged liposomes, both in the reticuloendothelial cells and in the hepatocytes. A striking difference exists, nevertheless, in the appearance of the two types of cells, 1 hour and 7 1/2 hours after injection. The specific antibodies never inhibited more than 50% of the activity of liver acid alpha-glucosidase, and there was no significant modification in the glycogen content of this organ. This work points to the influence of the composition of liposomes, in determining the entrapment of proteins, the toxicity, the uptake by the liver, and the distribution of these lipidic spherules in the different types of cells. This factor will have to be taken into account if liposomes are to be used as vectors in the treatment of lysosomal storage diseases.


Use of coercion in the outpatient treatment of alcoholism. Attendance patterns of voluntary patients at an alcoholism clinic were compared with those of persons convicted of drunken driving who were required to attend the clinic as a condition of probation. The attendance rate of the coerced patients was much superior to that of the voluntary patients.


A randomized pilot study comparing two regimens in the treatment of squamous cell carcinoma. Thirty-five patients with epithelial carcinomas not considered to be surgically resectable were randomized to receive two different chemotherapy regimens. Regimen 1 was CCNU followed by bleomycin, and Regimen 2 was a combination of CCNU, bleomycin, methotrexate, and vinblastine. Five of 14 patients treated with CCNU-bleomycin had partial responses. Three of 15 patients treated with the four-drug combination had a partial response. The toxicity of the four-drug regimen was significantly greater than that of the two-drug regimen, while the response rate was greater among those patients treated with two drugs. No significant clinical infections occurred despite the fact that leukopenia and thrombocytopenia were more severe and frequent with the four-drug regimen. Most importantly, the two-drug regimen is well tolerated as an outpatient procedure. Two of the two-drug and one of the four-drug recipients were converted from an inoperable to an operable state. In view of the fact that there has not heretofore been an effective chemotherapeutic regimen for Stage III and IV squamous carcinoma, this is a significant observation. It is of note that patients with squamous cell carcinoma of the head and neck were those who responded best to this treatment, although the treatment is worthy of consideration for advanced squamous carcinoma in other areas.


Ascending aorta-right pulmonary artery shunt. Seventy-seven patients with ascending aorta-right pulmonary artery shunt were reviewed; 48 had tetralogy of Fallot, 9 had pulmonary atresia, 11 had transposition of the great vessels with pulmonary stenosis, 4 had tricuspid atresia, and 5 had miscellaneous complex lesions. Their ages ranged from one day to 13 years. The over-all mortality rate was 17.8 per cent. Ten patients underwent total repair and takedown of the shunt with no deaths. The problems of increased pulmonary flow, pulmonary hypertension, preferential flow to one lung, kinking and distortion of the pulmonary artery, and the technical difficulties at the time of takedown were reviewed and discussed.


Right atrial partition and right ventricular exclusion: another surgical approach for complex cyanotic congenital heart disease. A 5-year-old child with asplenia, situs inversus, single ventricle, common atrium, severe subvalvular pulmonary stenosis, 1-transposition of the great arteries, and absent inferior vena cava presented with severe limitation (resting arterial saturation 74 per cent). At operation, the systemic venous atrium was partitioned with a Dacron baffle, so that hepatic venous and coronary sinus blood was enabled to drain with the pulmonary venous blood into the single ventricle and aorta. The superior portion of this atrium was anastomosed to the divided main pulmonary artery, so that most of the systemic venous blood was allowed to flow directly to the lungs. Intraoperative hemodynamic studies revealed a pulmonary artery pressure of 12/9 mm. Hg and a superior vena caval flow that was 88 per cent of the ascending aortic blood flow. Follow-up catheterization studies revealed an intact partition, no anastomotic gradient, superior vena cava pressure of 20 mm. Hg, arterial saturation of 84 per cent, and excellent flow of contrast from superior vena cava to atrium to pulmonary artery. Exercise tolerance was markedly improved, and chronic fluid retention was not observed. This operation offers a new alternative for long-term palliation of complex lesions amenable to exclusion of the right ventricle, such as single or common ventricle with unreconstructable anomalies of the atrioventricular valves.


Levamisole in advanced human breast cancer. A clinical trial of levamisole, an orally effective modifier of the immune response, is reported in women with primary inoperable breast cancer (stage III). After being rendered clinically disease-free by radiotherapy to the breast, supraclavicular area, and axilla, patients were allocated alternately to a control group (no further treatment) and a levamisole-treated group (150 mg orally three times a week on alternate weeks) and were followed-up by physical examination and laboratory tests. In 43 patients (23 control and 20 levamisole), there was significant prolongation of the median disease-free interval (25 v. 9 months) and survival (90% v. 35% alive at 30 months) in the levamisole-treated group compared with the controls. Levamisole treatment was also associated with an increase in the percentage and intensity of delayed hypersensitivity skin reactions and in the absolute lymphocyte-counts. No significant toxicity of levamisole was observed.


Failure of somatostatin to correct manifest diabetic ketoacidosis. Juvenile diabetic patients were studied 60-72 hours after insulin withdrawal when moderate ketoacidosis had developed. Somatostatin infusion for 4 hours in five patients resulted in almost complete suppression of plasma pancreatic glucagon and growth hormone, a fall in plasma-cyclic-adenosine-monophosphate (A.M.P.) concentrations, and a large fall in plasma-glucose concentration. After infusion plasma concentrations of these substances rose again. Blood-ketone-bodies, plasma-free-fatty-acids (F.F.A.), and plasma glycerol concentrations, however, did not decrease appreciably with somatostatin administration. In three patients 2 to 3 h somatostatin infusions were twice superimposed upon a continuous 9-5 h insulin infusion (1 unit/h). An insulin effect was noticeable within 30 minutes, with pronounced falls in the concentrations of plasma glucose, pancreatic glucagon, F.F.A., and blood-ketone-bodies. There was no significant change in these patterns when somatostatin was administered or withdrawn. These results do not indicate that somatostatin infusion would be useful in the treatment of manifest diabetic ketoacidosis.


Pancreatectomised man: A model for diabetes without glucagon. The proposition that glucagon plays an essential part in maintaining hyperglycaemia in diabetes has been investigated by the study of 5 totally pancreatectomised subjects and 5 age and sex matched insulin-dependent diabetic patients. True basal glucagon values were obtained by the use of a new affinity chromatography technique. The mean fasting plasma-glucose levels of the pancreatectomised subjects was 251 +/- 46 mg/dl. The mean fasting plasma-glucagon level was not significantly elevated above zero (1-3 +/- 0-6 pmol/l) and showed no change following arginine. In the 5 insulin-dependent diabetics the mean fasting plasma-glucagon level of 17-2 +/- 5-3 pmol/l rose to a maximum at 25 minutes of 103-6 +/- 27-5 pmol/l during infusion of arginine. These findings imply the absence of a significant number of normally functioning alpha cells in extrapancreatic sites in man and demonstrate that pronounced hyperglycaemia may occur in the absence of glucagon. Glucagon is probably not of primary importance in the hyperglycaemia of insulin-dependent diabetics.


Treatment of attacks in hyperkalaemic familial periodic paralysis by inhalation of salbutamol. In fifteen patients with hyperkalaemic familial periodic paralysis, inhalation of salbutamol alleviated hyperkaleamia and paralysis precipitated by exercise or oral administration of potassium chloride. In-vitro studies with rat soleus muscles indicated that the hypokalaemic effect of salbutamol is related to stimulation of the active coupled transport of sodium and potassium in muscle cells. Follow-up studies proved that the inhalation of salbutamol is a simple and adequate method for the treatment of the paralytic episodes in these patients.


Transcendental meditation in hypertension. Individual response patterns. Seven selected hypertensive patients were stabilized on drugs at a research clinic. Subjects learned transcendental meditation (T.M.), were seen weekly, and took their own blood pressure several times daily. After 12 weeks of T.M. six subjects showed psychological changes and reduced anxiety scores. Six subjects also showed significant reductions in home and four in clinic blood-pressures. Six months later four subjects continued to derive psychological benefit and two showed significant blood-pressure reductions attributable to T.M. at home and clinic.


Autoantibodies to Tamm-Horsfall protein, a tool for diagnosing the level of urinary-tract infection. In sera from ten girls with acute pyelonephritis, antibodies of the IgG-class to Tamm-Horsfall protein were found in significantly increased titres; in contrast, titres in sera from six girls with acute cystitis did not differ from those of healthy controls. The results suggest that determination of Tamm-Horsfall antibodies may prove useful for differentiating between upper and lower urinary-tract infection.


Hyperprolactinaemia and luteal insufficiency. Prolactin interferes with the function of the corpus luteum, as was demonstrated by repeatedly finding a short luteal phase in the ovulatory cycles of two hyperprolactinaemic women after prolactin supression by bromocriptine had been discontinued. In hyperprolactinaemic ovulatory cycles excretion of pregnanediol in urine was less and plasma-progesterone concentrations were lower than in normo-prolactinaemic cycles. When the amenorrhoea-galactorrhoea syndrome recurred the symptoms appeared in the following order: (1) serum-prolactin increased and the luteal phase became shorter; (2) galactorrhoea appeared; (3) ovulation was missed; (4) menstruation was missed. All these abnormalities may be treated by prolactin suppression, but the effect of treatment does not persist for long after bromocriptine administration ceases.


HLA-B27 and modified bone formation.

HLA-B27 and modified bone formation. Of the many associations between histocompatibility antigens and human diseases a prominent one is that between HLA-B27 and inflammatory arthropathies. Hypotheses to explain this association include the B27 gene being linked to a specific immune-response gene required for disease expression and the B27 antigen acting via molecular mimicry with a microorganism or as a microorganism receptor. Alternatively, the HLA-B27 gene might be closely related to a gene which influences bone formation. The finding of a significant association between B27 and Forestier's disease, a disease characterised by abundant new bone formation, supports such a hypothesis.


Evaluation of radiography and isotopic scintigraphy for detecting skeletal metastases in breast cancer. Experience with technetium-phosphate compounds for skeletal scintigraphy in patients with breast cancer was analysed. When tumours were 5 cm or less in diameter (T1-2N0-1M0) metastases were demonstrated by radiographs in 1-7% (2/114). However, when radiography did not demonstrate metastases, lesions were found by scintigraphy in 41-3% (19/46). When lesions demonstrated by scintigraphy at the same site as abnormalities regarded as "benign" by radiography were excluded, 23% (11/46) had scintigraphs strongly suggestive of metastases. It is proposed that routine radiographic skeletal survey for patients presenting with breast cancer be abandoned, and replaced by skeletal scintigraphy, chest radiography, and specific localised radiographs of lesions demonstrated by scintigraphy. It is suggested that with this policy the development of expertise in interpreting scintigraphs will be accelerated, the cost of pre-treatment assessment will be reduced, and clinical management rationalised.


Therapy with parent's lymphocyte transfer factor in children with infection and malnutrition. Transfer factor (T.F.) prepared from 5 x 10(8) lymphoid cells from 500 ml of a parent's blood was given to 40 Australian aboriginal children aged 2-46 months who had been in hospital with acute infection. Many had protein-calorie malnutrition. These and a control group of 35 similar children were assessed blind for at least 12 months. In T.F.-treated children there were significantly fewer episodes of diarrhoeal disease for periods in excess of 26 weeks. Recurrent moderate diarrhoeal disease was particularly reduced, and the onset of severe gastroenteritis may have been delayed. There was no protection against chest, middle-ear, or skin infection.


Apatite deposition disease. A new arthropathy. A method for identifying particles of crystalline calcium hydroxyapatite in synovial fluids and biopsy material has been developed with high-resolution scanning electron microscopy and an energy-dispersive micro-analytical system. Particles of hydroxyapatite were identified in the joints of six patients diagnosed as having osteoarthritis, three of whom had acute inflammatory episodes with effusions into the joints. Apatite was not identified in joints affected by rheumatoid arthritis and other types of arthropathy. Animal studies showed that hydroxyapatic crystals can cause an acute inflammatory reaction, and this has been confirmed by experimental studies in man. It is suggested that a third type of crystal-deposition disease should be recognised--namely, calcium-hydroxyapatite crystal-deposition disease.


Enzymic analysis of cardiac biopsy material from patients with valvular heart-disease. Cardiac tissue obtained by left-ventricular endomyocardial biopsy from patients with valvular heart-disease was assayed for marker enzyme activities of subcellular organelles and these were correlated with left ventricular function as assessed by haemodynamic studies. In patients with poor left ventricular function, calcium-dependent adenosine-triphosphatase (A.T.P.ase) activity, predominantly localised to the myofibrils, was strikingly reduced. Activity of lactate dehydrongenase, a cytosol enzyme, was significantly increased in tissue from patients with poor left ventricular function. The activity of enzymes associated with sarcolemma (5'-nucleotidase), mitochondria (glutamate dehydrogenase and monoamine oxidase), microsomes (neutral alpha-glucosidase), and lysosomes (acid phosphatase, N-acetyl-beta-glucosaminidase) was no different in patients with good or poor left ventricular function. It is suggested that reduced myofibrillary A.T.P.ase concentration is the biochemical basis for the impaired ventricular function.


A double-blind trial of the effect of wheat bran on symptoms of irritable bowel syndrome. 59 outpatients with irritable bowel syndrome participated in a randomised double-blind trial. The patients in the treatment group received three biscuits daily each containing 10 g of ordinary miller's bran, whereas the patients in the control group received wheat biscuits of a similar appearance. The treatment period was 6 weeks. 52% of the patients in the treatment group noted subjective improvement compared with 65% in the control group. The results of this trial do not support the routine use of miller's bran in irritable bowel syndrome.


Anticonvulsants and parental epilepsy in the development of birth defects. The results of two studies, one in Finland and one in the U.S.A., raise the possibility that fetal damage previously attributed to phenytoin and other anticonvulsant drugs, principally phenobarbitone, may be due to epilepsy itself. In the U.S.A., drug-exposure information was collected before delivery in a cohort of 50 282 mother/child pairs. The total malformation rate in 305 children born to epileptic mothers was 10.5%, as against 6.4% in the remainder (p less than 0.01); corresponding rates for major malformations were 6.6% and 2.7%. When the fathers had epilepsy, the malformation-rates in their children were intermediate. The rates did not vary significantly according to maternal anticonvulsants therapy. Mental and motor scores as 8 months of age, and intelligence quotient scores at 4 years were lower in children of epileptic mothers, but not in children of epileptic fathers. The scores showed only random variation according to maternal anticonvulsant therapy. In Finland, 2784 children with craniofacial anomalies were compared with an equal number of normal children; 8 and 2 mothers, respectively, received anticonvulsants, while pregnant, for epilepsy. In that study, the separate effects of the disease and its treatmet could not be evaluated. Both studies did not find evidence of fetal damage when phenobarbitone was taken for indications other than epilepsy.


Abnormalities in the apparently normal bowel mucosa in Crohn's disease. The uninvolved intestinal mucosa away from the affected areas in Crohn's disease has always been considered to be macroscopically and microscopically normal. Rectal biopsy specimens from 13 patients who had had Crohn's, disease elsewhere in the bowel, but never in the rectum, showed significant increases in the plasma-cell density in the lamina propria, in the volume of the lamina propria, and in the glucosamine-synthetase activity of the specimens, compared with a control series of patients with the irritable-colon syndrome. It is suggested that the colonic mucosa is always abnormal in Crohn's disease even if macroscopical and histological examination shows an apparently normal mucosa.


Cerebral thrombosis, cerebral haemorrhage, and ABO blood-groups. The ABO blood-group distributions of 1460 patients who had died from a stroke were compared with those of a control group of 20 705 controls selected at random from the healthy population at risk (i.e., over thirty-five years of age and matched for age and sex ratio). The cause of death was certified as cerebral thrombosis in 329 cases and as cerebral haemorrhage in 482 cases, these diagnoses being established in neurological hospitals; the remaining 649 cases had an unspecified type of stroke, the diagnosis being made by general practitioners. In the group with unspecified type of stroke the blood-group distribution was practically the same as the distribution in the controls. In the thrombosis cases there was an excess of blood-groups A and AB and a deficiency of O and B; in cerebral haemorrhage this situation was reversed. However, these were only trends; the differences were not significant at the 5% level. A statistically significant difference did emerge when the A+AB excess in thrombosis was contrasted with the O+B excess in haemorrhage, suggesting that this difference might be accounted for the major A subgroup (A1) and, consequently, A1B.