Fractionation of mouse alpha-fetoprotein. 4 distinct alpha-fetoprotein (AFP) containing fractions were obtained upon ion-exchange chromatography of late-gestational fetal mouse extracts. Despite this chromatographic heterogeneity, the individual AFP isolates were antigenically indistinguishable.


Autoradiography and differential hemoglobin staining as aids to the study of fish hematology. The confused state of the existing knowledge regarding the cell types in peripheral blood and hematopoiesis of fishes has been highlighted. Inadequacies of techniques presently used have been pointed out and the advantages of using Graham-Knoll's method for haemoglobin staining when counter stained with Giemsa together with autoradiography using tritiated thymidine have been demonstrated.


Plasma steroid and protein hormone concentrations in patients with benign prostatic hypertrophy and in normal men. Serum levels of testosterone, FSH, LH and prolactin were measured by RIA in patients with BPH (benign prostatic hypertrophy) and in control men. The testosterone concentrations in the BPH group were significantly lower than that of the control group. Serum concentrations of FSH, LH, prolactin and estradiol, did not differ significantly in the 2 groups.


Macrophage immunity to influenza virus: in vitro and in vivo studies. Using M-TUR, a macrophage-adapted avian influenza A virus (Hav1, Nav3), antiviral resistance of peritoneal macrophages obtained from specifically or nonspecifically immunized mice towards in vitro infection was assessed. M-TUR grew to high titers in macrophages from nonimmune mice thereby causing a marked cytopathic effect. In contrast, peritoneal macrophages from mice specifically immunized with TUR virus were not affected by infection with M-TUR in vitro. This antiviral immunity was specific: mice immunized with antigenetically unrelated influenza strains such as influenza A/Hong Kong/1/68 (H3, N2) or influenza B/Lee yielded susceptible macrophages. Specific macrophage immunity could be abrogated by trypsin treatment in vitro. Susceptible macrophages from nonimmune hosts became resistant following in vitro exposure to homologous anti-TUR sera. Peritoneal exudate cells from BCG-infected animals were less susceptible to in vitro challenge with M-TUR than control macrophages. In vivo treatment of mice with the unspecific immunostimulants BCG or Corynebacterium parvum did not protect the animals against lethal infection with a hepatotropic variant of TUR.


Cyclic AMP and lipogenesis in fat cells from thyroidectomized rats. Thyroid hormones regulate lipid metabolism by affecting lipogenesis as well as lipolysis. The present paper discusses the way thyroidectomy induced an enhancement in lipogenesis in rat fat cells. The doubling in the conversion of glucose to CO2 and fatty acids seen after thyroidectomy was found to be due to a modification in the actual pathway of glucose metabolism: there was a preferential stimulation of the conversion of glucose to CO2 by the pentose cycle (utilisation of [1-14C]glucose) while the production of fatty acids and glyceride-glycerol proceeded, respectively, much more, or only slightly more, via the pathway of [6-14C]glucose metabolism. Studies employing the phosphodiesterase inhibitor MIX, or the cyclic AMP analogue, DBcAMP showed that the lipogenic process depends on cyclic AMP. As the stimulatory effect of thyroidectomy was not abolished, however, lipogenesis must be under the independent control of both cyclic AMP and absence of thyroid hormones. Insulin, a further mediator of lipogenesis was found to further enhance the already preexisting high conversion of glucose to CO2 in fat cells from thyroidectomized rats. It is concluded that at least three factors modify lipogenesis: thyroidectomy, cyclic AMP and insulin; each achieving its effect in an independent manner.


The production and control of anti--venous sera. This paper describes the species of venomous snakes found in Costa Rica as well as anti-venom sera prepared by the Instituto Clodomiro Picado: polyvalent sera for use in human or veterinary medicine, and anti-Micrurus and anti-Lachesis monovalent sera. These sera are prepared in horses, purified with ammonium sulphate, then concentrated and delivered either in liquid or freeze-dried form. Controls are in accordance with NIH recommendations. For cases of hypersensitivity to horse serum, a polyvalent anti-Bothrops and anti-Crotalus serum from sheep has been prepared.


Program for the expansion and improvement of biologics production and control in Latin America. The production and control of biologicals sufficient to meet the needs of the countries in the Americas has been of basic concern to the Pan American Health Organization for many years. This is evidenced by the Ten-Year Health Plan for the Americas, drawn up in Santiago, Chile in 1972 by the Health Ministers of the Americas, in which the goal of making the Region self-sufficient in all of the essential immunizing agents within a ten-year period, was established. At present only Canada and the United States are more or less self-sufficient in this regard. All of the others are, to a greater or lesser degree, dependent on imports. In PAHO's present program the greatest emphasis is being placed on the development of effective control systems, particularly at the national level. The steps taken to achieve this goal and the progress to date are outlined.


[Determination of antigen structures of Pasteurella multocida: the preparation of vaccines]. The discovery of the role played by somatic antigens of the Pasteurella multocida A and D capsular types in the immunization of susceptible species makes it necessary to type the somatic antigens of the isolated strains prior to any action to induce active or passive immunity. Only immunoprecipitation in gel was chosen out of the various typing techniques proposed, and a correlation was established between the capsular types (according to Carter), somatic types (according to Namioka) and types determined by immunoprecipitation in gel (according to Heddleston). Parallel to this, the same technique was developed for strains of Pasteurella multocida B and E capsular types, and the immunogenicity of these capsular antigens was proven.


Demonstration of a novel cellular retinol-binding protein, F-type, in hepatocellular carcinoma. Vitamin A level and the cytosol-binding proteins specific for vitamin A ere studied in human tumor and its surrounding tissue. The tissues examined were 10 hepatocellular carcinomas which were surgically removed, 4 other malignant tumors (2 metastatic liver cancer and one each of gastric cancer and glioma), and 3 human fetal livers. Compared with surrounding tissues, considerable decrease of vitamin A content was observed in the hepatocellular carcinoma suggesting local deficient state of the vitamin. In addition to cellular retinol-binding protein (CRBP) and retinoic acid-binding protein (CRABP), a new molecular species having affinity for both retinol and retinoic acid was detected in the cytosols obtained from hepatocellular carcinoma as well as glioma by means of gel filtration on Sephadex G-75. With regard to ligand specificity, the protein was found to be similar to cellular retinol-binding protein, F-type or CRBP(F) which was originally recognized in the fish eye cytosol. Since the protein was also demonstrated in human fetal liver, CRBP(F) is considered to be an oncofetal protein in nature. The present study further revealed that CRBP(F) was detected in 80% of hepatocellular carcinoma (whereas plasma alpha-fetoprotein was significantly elevated only in 50%), and hepatocellular carcinoma contained CRBP(F) in a larger amount than CRABP.


HLA and cell-mediated immunity in HBsAg negative chronic active hepatitis. In 20 patients with severe HBsAg negative chronic active hepatitis a significant association has been found between the HLA determinants A1, B8, and Dw3 and cell-mediated immunity directed against human liver specific lipoprotein in vitro. This observation induces further speculation that genetic immunoregulatory action might be involved in the pathogenetic mechanisms of chronic active hepatitis in vivo.


[Experimental yolk sac tumors in the rat (author's transl)]. A study on the experimentally induced yolk sac tumor in the rat was made in special regard to the characteristics and origin of tumor cells. Pregnant rats which fetuses were removed on the 12th day of gestation, developed tumors derived from the fetal membranes left outside the uterus, which were composed of differentiated teratomas and yolk sac tumors. Serial observation of the oncogenesis revealed that an early lesion of the yolk sac tumor appeared in a nodule found as early as 3 weeks after the fetectomy and production of alpha-phetoprotein (AFP) was observed histochemically in the tumor cells 5 weeks after the fetectomy. A cultured cell line established after cloning from the transplantable yolk sac tumor which had been induced similarly and converted into ascitic form was also investigated. Light and electron microscopic studies on both induced tumors and the cultured cells indicated a similarity of AFP producing tumor cells with parietal yolk sac cells and of PAS-positive hyaline-like substance with the Reichert membrane. It is reasonable to conclude that the yolk sac tumor observed is regarded as a parietal yolk sac carcinoma described by Pierce. Histogenesis of the tumor is also discussed.


Trasylol prevents trypsin-induced shock in dogs. The effect of simultaneous intravenous administration in the dog of bovine trypsin and Trasylol followed by continued infusion of Trasylol was studied. Special attention was paid to the interchange between the dominating plasma protease inhibitors alpha1-antitrypsin and a-macroglobulins and to the disappearance of Trasylol and its trypsin complexes from the circulation. The following results were obtained: 1) Trypsin was preferentially bound by the alpha-macroglobulins, though Trasylol is a strong trypsin inhibitor. 2) On saturation of the alpha-macroglobulins, a considerable amount of trypsin was bound by alpha1-antitrypsin. 3) Trasylol was bound to the trypsin-alpha-macroglobulin complexes and then rapidly eliminated from the circulation. 4) On saturation of the alpha-macroglobulins, Trasylol was identified in a free form but increasing amounts of Trasylol were also bound to trypsin. This could be explained not only by direct complexation of Trasylol and trypsin but also by a transfer of trypsin from unstable trypsin-alpha1-antitrypsin complexes to free Trasylol.


Observations on the results of specific histochemical techniques and empirical staining methods on several tissue/organ types using a variety of fixing fluids. Several histotechniques (Harris hematoxylin and alcoholic eosin, Gomori's reticulum method, Verhoeff's elastic stain; Gomori's trichrome, Mallory's aniline blue collagen stain and the Cameron and Steele method) were each used with a number of fixatives (Lavdowsky's chromic acid, Karnovsky's glutaraldehyde/paraformaldehyde, neutral buffered formalin, Lavdowsky's formalinacetic acid-alcohol, Zenker's and Helley's). Lavdowsky's chromic acid fixative provided superior results as compared to the other fixing fluids and its use was critical with respect to the trichrome or connective tissue staining procedures. The Cameron and Steele modification used with Lavdowsky's chromic acid fixative gave the best differentiation of tissue types and was also the most reproducible.


Duodenal glucagonoma: a case report.

Duodenal glucagonoma: a case report. A case of a primary carcinoid islet cell tumor of the duodenum is reported, demonstrated by histochemistry, electron microscopy, and immunofluorescence to be composed of alpha cells containing glucagon-like material. The patient was found on admission to have hyperglycemia and a diffuse skin rash. Primary duodenal glucagonoma has not been previously reported.


Effects of alkylating agents on lymphocytes from controls and from patients with Fanconi's anemia. Studies of sister chromatid exchanges, chromosome aberrations, and kinetics of cell division. The frequency of sister chromatid exchanges (SCE) and chromosome aberrations and the dynamics of cell division in peripheral blood lymphocytes of four patients with Fanconi's anemia were studied after in vitro exposure to alkylating agents TEPA and mitomycin. SCE frequency was significantly increased even after very low doses of mutagens, while chromosome aberrations were significantly increased only after high doses (0.160 micrograms/ml mitomycin and 10(-5) M TEPA). The responses of Fanconi's anemia cells and control cells did not differ significantly. The increased frequency of both SCE and chromosome aberrations was accompanied by gradual delay of cell division, which was most conspicuous in cells from patients with Fanconi's anemia.


Complete deficiency of constitutive heterochromatin on a human chromosome 9. In cultured amniotic fluid cells a mediocentric chromosome 9 appeared to be completely deficient in constitutive heterochromatin when stained with distamycin A and DAPI. In addition, this deficient chromosome was found in a blood cell culture from the father. Both the father and the child after birth were phenotypically normal. Evidently, a considerable heterozygotic deficit of chromosome 9 heterochromatin can be tolerated without affecting the phenotype. The heterochromatin defect was also shown by G11-staining. Distamycin A-DAPI staining is highly reproducible and is recommended as a fluorescent alternative to often less successful G11-methods for the detection of heteromorphism of chromosome 9.


Group-specific component (Gc) and transferrin (Tf) subtypes ascertained by isoelectric focusing. A simple nonimmunological staining procedure for Gc. Simultaneous subtyping of Gc and TfC by isoelectric focusing allows us to compute the following gene frequencies for the Belgian population: Gc1S = 0.543 Gc1F = 0.167 Gc2 = 0.290 TfC1 = 0.784 TfC2 = 0.206 TfB = 0.007 TfD = 0.003. The Gc bands were precipitated by sulfosalicylic acid instead of by immunofixation.


Determinant selection and macrophage function in genetic control of the immune response. The immune response to insulin, in both mouse and guinea pig, is under control of H-linked immune response genes. When immunized with either pork or beef insulin in CFA, both strain 2 and 13 guinea pigs respond by antigen-specific lymphocyte proliferation and synthesis of specific antibody. The specificities of the elicited antibodies and indistinguishable between these inbred strains. By constrast, strain 2 T cells recognized a distinct region of the A chain alpha loop consisting of amino acid residues 8, 9 and 10, while strain 13 T cells see an as yet undefined region of the B chain. H2b (A chain alpha loop responder) and H2d (B chain responder) mice similarly discriminate which areas of the molecule are recognized by their T lymphocytes. The function of the Ir gene in both the guinea pig and mouse appears to be an intramolecular selection of discrete regions within the antigen for recognition by the T cell. The data presented suggest that this function operates at the level of the macrophage.


The cell-mediated immune response of the pig to orally administered antigen. The immune response of pigs to oral immunization with dinitrophenylated bovine gamma globulin (DNP-BGG) was assessed using the indirect macrophage migration assay. Two days' exposure to the antigen resulted in the inhibition of macrophage migration by supernatants of cultures from the intestine but not the mesenteric lymph node or spleen. Stimulation for 7 days resulted in macrophage inhibition by supernatants of lymphocyte cultures prepared from the intestine, mesenteric lymph node and spleen. Supernatants of splenic lymphocytes from animals stimulated for 2 or 3 days produced enhancement of macrophage migration.


Immunological unresponsiveness in mice. II. Cellular basis of immunological unresponsiveness induced in foetal and neonatal mice by transfer of human gamma-globulin by the maternal route. The cellular basis of the mechanism of immunological tolerance to human gamma-globulin (H gamma G) induced in foetal and neonatal mice by materno-foetal or materno-neonatal transfer after a single injection of tolerogen (deaggregated H gamma G) into the mothers was investigated using a cell transfer system and assays of passive haemagglutinating antibodies and plaque-forming cells to H gamma G. The results demonstrated that B cells are mainly involved in the tolerance induced on the fourteenth day of gestation, whereas inactivation of T cells may account for the tolerance induced on the eighteenth day of gestation and in the neonatal stage. Treatment of the mothers with tolerogen and then anti-H gamma G serum reduced the tolerance induced on the fourteenth day of gestation, but did not affect that induced on the eighteenth day of gestation and in the neonatal stage. Cell transfer experiments showed that B-cell tolerance induced on the fourteenth day of gestation was prevented by passive antibody, while T-cell tolerance induced on the eighteenth day of gestation and in the neonatal stage was not affected by passive antibody. Assay of the anti-DNP antibody response after immunization with DNP10-H gamma G showed that treatment of mice with the tolerogen on the eighteenth day of gestation, but not the fourteenth day of gestation, inactivated H gamma G-reactive helper cells. The significance of these results is discussed in relation to the results of the cell transfer experiments described as above.


Studies on the immune response in chickens. III. Effect of substitution of carrier on elicitation of anti-hapten antibody responses and generation of hapten-specific memory. The role of carrier in both elicitation of anti-hapten antibody responses and generation of hapten-specific memory in chickens were studied using dinitrophenyl (DNP) conjugate of T-independent antigen carrier (DNP--Ficoll), that of isologous serum protein DNP--chicken serum albumin (CSA), and heavily conjugated DNP--bovine gammaglobulin (BGG). All of these DNP--conjugates could generate DNP specific memory as markedly as moderately conjugated DNP--BGG. Both DNP--Ficoll and DNP--CSA hardly elicited the primary and secondary anti-DNP antibody responses. Heavily conjugated DNP--BGG did not elicit a significant primary anti-DNP response as moderately conjugated DNP--BGG. It is suggested therefore that, when chickens are stimulated with hapten-carrier conjugates, hapten-specific memory is generated independently of T-cell function, but the elicitation of both the primary and secondary anti-hapten antibody responses is somehow relevant to T-cell function, although the T-cell dependence is less marked in the secondary than in the primary response.


Studies on the resistance to tolerance induction against human IgG in DDD mice. II. Tolerogen-resistant T-cell population in the spleen. Further studies were carried out to investigate cellular sites of the resistance to the induction of immunological tolerance to HGG in DDD mice, assuming the presence of a subset of tolerogen-resistant splenic T cells. Spleen-seeking T cells were shown to be relatively resistant in comparison with lymph node-seeking T cells. The existence of a tolerogen-resistant T-cell subpopulation was indicated from the experiments demonstrating that tolerance was easily attained after adult thymectomy and that lymph node T cells became much more resistant to tolerance induction after adult splenectomy. The latter experimental result might also exclude the possibility of differences in microenvironment (probably in A cells) between spleen and lymph node. An attempt was made to investigate a possible involvement of A cells in the induction of tolerance. A cells were deprived in vivo by irradiation of the host 3 days prior to spleen cell transfer and in vitro by passing a spleen cell suspension through a Sephadox G-10 column. The deprivation of A cells resulted in priming of the host by the tolerogen rather than easier tolerance induction. No suppressive activity was observed in lymph node cells from tolerized mice. These results suggest that there exists a set of T cells, generated relatively recently in the thymus, preferentially migrating into spleen and there becoming resistant to tolerance induction.


Chemotactic factor receptors of human PMN leucocytes. I. Effects on migration of labelling plasma membrane determinants with impermeant covalent reagents and inhibition of labelling by chemotactic factors. The mechanism of stimulation of human PMN leucocyte-directed migration by chemotactic factors was studied by pre-labelling plasma membrane determinants with impermeant covalent reagents and assessing the effects of such modification on spontaneous mitration and chemotaxis in modified Boyden chambers. Pre-treatment of PMN leucocytes with 10(-9) to 10(-6) M isethionyl acetimidate, which selectively labels amino groups, enhanced spontaneous migration and concomitantly inhibited chemotaxis to fragments of the fifth component of complement (C5fr), 12-L-OH-5,8,10,14-eicosatetraenoic acid (HETE) and several formyl--methionyl (f--Met) peptides to an extent that was inversely related to the magnitude of the chemotactic response of untreated PMN leucocytes. Para-chloromercuribenzene sulphonate, which selectively labels sulphydryl groups, inhibited chemotaxis to diverse stimuli without substantially influencing spontaneous migration, while the diazonium salt of sulphanilic acid, which labels several types of plasma membrane determinants, altered neither spontaneous nor chemotactic migration. Incubation of PMN leucocytes with various concentrations of [3H]-isethionyl acetimidate labelled from 33,000 amino groups per PMN leucocyte at 10(-6) M to over 800,000 at 10-4) M, a reaction that was substantially inhibited by chemotactic concentrations of C5fr and HETE, but not by f--Met peptides. Subcellular fractionation of PMN leucocytes labelled with [3H]-isethionyl acetimidate localized the radioactivity to membrane-rich fractions. Free amino groups thus appear to be functionally critical determinants of some chemotactic factor receptors on the plasma membrane of PMN leucocytes.


Autoregulation of germ tube formation by Candida albicans. Germ tube formation by Candida albicans is at least partially controlled by a product(s) of the yeast phase of the organism which is released from cells upon incubation at 37 degrees C in tissue culture medium or fetal calf serum. This germination regulatory substance is stable under conditions of lyophilization and heating of 70 degrees C, but becomes inactivated at pH values of 4.0 and 9.5. A germination regulatory substance was produced by both strains of C. albicans tested and by a strain of C. tropicalis. Production does not appear to be a universal characteristic of yeasts because the factor could not be recovered from either Cryptococcus laurentii or Candida parapsilosis. Previously described C. albicans germination inhibitors such as cysteine, tryptophol, and phenylethyl alcohol appear not to be the substance described here. Because of the ability of the factor to influence C. albicans morphology, we have designated it morphogenic autoregulatory substance.


Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. The heat-labile enterotoxin (LT) has been isolated in homogeneous form with high specific activity from three sources: cell-free supernatant, NaCl extract, and whole-cell lysates of an enterotoxigenic Escherichia coli strain. In vitro immunological assays were used in lieu of tedious and highly variable bioassays to recognize fractions with activity. This revealed that the major portion of the LT remained adherent to columns containing agarose, from which it could be eluted quantitatively in practically homogeneous form by galactose. Isolated LT has remarkable similarities to the cholera enterotoxin (choleragen) in both subunit structure and amino acid composition, although there are also notable differences in these two enterotoxins, which are related immunologically and by mode of action. Unlike choleragen, in which the A region is totally nicked, E. coli LT, depending on its source, is activated by proteolytic processing. The activity of LT is equivalent to that of choleragen in bioassays on adrenal cells, in rabbit skin, and in rabbit ileal loops, especially when, depending on the source of material, the LT has been activated by treatment with trypsin. The whole-cell lysate is the richest source of LT.


Antigenic specificity of neutralizing antibody to cholera toxin. Selected rabbit antisera to cholera toxin antigens and convalescent cholera patient sera were analyzed using the permeability factor neutralization test and two sensitive in vitro serological assays specific for cholera toxin, cholera toxin A subunit, and cholera toxin B subunit. The results indicated that antisera to cholera toxin contained toxin-neutralizing activity as well as antibodies specific for both the A subunit and B subunit. It was clearly established that antisera to B subunit, devoid of significant anti-A subunit activity, neutralized the vascular permeability activity of cholera toxin. Antisera to A subunit contained neutralizing antibodies and antibodies to both A and B subunits. Absorption with B subunit removed both the toxin-neutralizing and anti-B subunit activities, while the anti-A activity was unaffected. Neutralizing antibody titers of rabbits immunized with B subunit were also observed to be significantly higher than neutralizing antibody titers of sera from A subunit-immunized rabbits, despite the overall similarity in anti-B subunit titers as determined by passive hemagglutination and radioimmunoassay of sera from the two groups of rabbits. Anti-alpha chain sera neither neutralized cholera toxin nor possessed significant antitoxin or anti-B subunit titers as determined by passive hemagglutination and radioimmunoassay. The anti-alpha chain sera contained high levels of antibody specific for A subunit, which is consistent with the hypothesis that the alpha chain is part of the A subunit structure. In contrast, the gamma chain was not shown to be antigenic. Sera from convalescent cholera patients possessed toxin-neutralizing antibody as well as passive hemagglutination and radioimmunoassay antibody against both A and B subunits.


Antigenic determinants in influenza virus hemagglutinin. Three antigenic determinants were revealed in H3 hemagglutinin of influenza A viruses isolated from 1968 to 1975. One of them was common for all viruses, and two others specified differences between the viruses possessing H3 hemagglutinin.


Inactivators of fibroblast interferon found in human serum. Investigation of the inactivators of fibroblast interferon has shown that those substances could be identified in every specimen of serum taken from normal individuals. These inactivators were found to destroy a mean of 82% of the interferon antiviral activity beyond that caused by the effects of temperature alone. The inactivators exerted no effect at 4 degrees C but were found in a variety of animal as well as human sera. Although slight differences were found to exist in the extent of inactivation when comparing normal human serum to serum from patients (i) receiving chronic hemodialysis, (ii) with jaundice, or (iii) with underlying malignancies, the differences did not appear to be clinically significant. In exploring the nature of the substances, we found that both dialyzable and non-dialyzable inactivators exist, but the former appear to be the major factors responsible for the unwanted process.


Distribution of anionic groups at the cell surface of different Sporothrix schenckii cell types. The distribution of anionic groups at the cell surface of yeastlike forms, hyphae, and conidia of Sporothrix schenckii was studied by staining with colloidal iron hydroxide and cationized ferritin. By using colloidal iron hydroxide it was shown that the external cell wall layer of one strain (strain 1099.18) could be resolved into two reactive sublayers and that these layers were present in many but not all cells of the same population. In contrast, most cells of another strain (strain 1099.12) were stained by colloidal iron hydroxide, but only one reactive layer was seen. Acidic layers of the yeastlike forms of the two strains were much thicker than those of conidia and hyphae. By the cationized ferritin staining procedure it was observed that the acidic layers of yeast forms sloughed off of cells, probably due to cell-cell or cell-medium attrition in shaken submerged cultures or to a process by which the outer layers detach from cells as they are replaced by newly synthesized ones. The colloidal iron hydroxide- and cationized ferritin-reactive cell surface layers of S. schenckii correspond to the previously described (L. R. Travassos et al., Exp. Mycol. 1:293-305, 1977) concanavalin A-reactive peptidorhamnomannan complexes, and their reactivity is probably due to the presence of acidic amino acids of low pK values rather than to glucuronic acid units.


Antigens of Micropolyspora faeni strains. Exocellular antigens from Micropolyspora faeni and Micropolyspora rectivirgula were prepared by growing them in a synthetic broth culture at 50 degrees C for 1 week under continuous shaking. The broth was separated from the organisms by filtration and the filtrate was dialyzed and freeze-dried. The cross-reactivity of these antigens were studied by antigen-antibody crossed-immunoelectrophoresis and the presence of similar antigens in different strains by tandem crossed-immunoelectrophoresis using rabbit anti-M. faeni and anti-M. rectivirgula sera. The results indicate that a number of antigenic components are common to the different strains of M. faeni and M. rectivirgula, while each strain demonstrates several specific antigenic determinants. Of the 26 clearly demonstrable precipitin arcs formed by crossed-immunoelectrophoresis, only 1 was detected in all the strains while 2 more were detected frequently. This study indicates that M. rectivirgula and M. faeni strains are not distinguishable from each other by morphological, biochemical or immunological criteria.


Characterisation of the reaginic response to sulphonamides in mice. Reaginic responses were induced in immunised mice with either 4-sulphanilamidobenzoic acid (4-SABA) or sulphamethoxazole (SMX) coupled to chicken gamma-globulin (CGG). The former was coupled through the carboxylic group of benzoic acid and the latter through the diazo derivative of the sulphanilamide group. The specificity of the reaginic responses obtained was assessed in each case by inhibition of passive cutaneous anaphylaxis in the rat. Immunisation with 4-SABA-CGG resulted in antibodies which recognised the sulphanilamide group as immunodominant and, therefore, they cross-reacted strongly with other sulphonamides. In contrast, SMX-CGG-induced IgE antibodies directed primarily to the methoxazole end of the molecule and cross-reactivity with other sulphonamides could not be demonstrated. This model may be useful for a more comprehensive study of allergic reactions induced by sulphonamides.


Effects of drugs on the acute inflammation following intraperitoneal injection of antigen into actively sensitised rats. When actively sensitised rats were injected intraperitoneally with antigen, the local reaction that ensued can be divided into two phases: an immediate reaction characterised by histamine and SRS-A release with an associated extravasation of plasma proteins, and a later reaction involving infiltration of neutrophilic polymorphonuclear leucocytes. When the immediate reaction was modified by BRL 10833 (which inhibits histamine release from rat mast cells and reduces extravasation of plasma proteins), there was no reduction in neutrophil infiltration. FPL 55712, an SRS-A antagonist, also failed to inhibit neutrophil infiltration. The beta-adrenoreceptor stimulants isoprenaline and salbutamol reduced neutrophil infiltration. Isoprenaline inhibited the extravasation of plasma proteins when given before antigen, but even when administered to rats after antigen, when extravasation was complete, it still inhibited neutrophil infiltration. Propranolol reversed isoprenaline-induced inhibition of neutrophil infiltration.


CEA and non-specific cross-reacting antigen (NCA) in medullary carcinomas of the thyroid. An immunohistological study of five cases of medullary carcinoma of the thyroid was undertaken with monospecific antisera against CEA and NCA. CEA was present in different areas of the tumors: cell cytoplasm, cell membrane at its apical pole when the tumor had a pseudoglandular organization, deposits in the lumen of pseudoglands, and peri- and extra-cellular deposits. From these extra-cellular deposits, CEA could easily reach the circulation, thus causing rising serum levels up to high values. NCA was found in the same tumors and in the same localizations.


Epstein-Barr virus-induced membrane antigens: immunochemical characterization of Triton X-100 solubilized viral membrane antigens from EBV-superinfected Raji cells. In an attempt to qualitatively identify the membrane antigen (MA) complex induced by Epstein-Barr virus (EBV) infection of lymphoblastoid cells, superinfected Raji cells were surface labelled with 125I by the lactoperoxidase method and solubilized with Triton X-100, then the 125I-labelled membrane proteins were precipitated by sera containing high antibody titers to MA. Analysis of these immune precipitates on sodium dodecyl sulfate polyacrylamide gel eletrophoresis identified four major EBV-specific membrane proteins with molecular weights (mol. wt) of 280,000, 250,000, 170,000 and 90,000. Sera from patients with Burkitt's lymphoma (BL), nasopharyngeal carcinoma (NPC) and infectious mononucleosis (IM) and from EBV-infected disease-free individuals showed differential patterns of reactivity to these antigens with some sera only recognizing three or less of the antigens. The results from invesigations with these sera also indicated that these major proteins were not related to EBV-induced viral capsid antigens (VCA) or the virus-associated early antigen (EA) complexes as defined by immunofluorescence. Metabolic labelling of EBV-infected Raji cells with [14C]glucosamine, followed by Triton X-100 solubilization and radioimmune precipitation, identified the 280,000, 250,000 and 90,000 components as glycoproteins. The lactoperoxidase-labelled 170,000 molecular weight component was not significantly glycosylated and, therefore, could not be categorized as a glycoprotein on the basis of this study. In addition, a glycoprotein with a mol. wt of 130,000 was identified by this approach which also appeared to be specified by EBV. The results from these investigations, therefore, indicated that the EBV-induced MA complex was composed of four major glycoproteins and one nonglycosylated high mol. wt protein.


Differential inducibility of Epstein-Barr virus in cloned, non-producer Raji cells. Cells of the human lymphoblastoid non-producer line Raji were cloned in soft agar. Individual colonies were isolated and analyzed for their inducibility of the Epstein-Barr virus-associated early antigen (EA). The induction of EA by the tumor promoter TPA varied among the different cell clones. Clones with very high and very low inducibility of the resident Epstein-Barr virus genome were further analyzed. Constant differences in the inducibility of EA were observed after activation by tumor promoters, 5-iododeoxyuridine or antibodies to human IgM. Induction of EA synthesis by superinfection with Epstein-Barr virus from the P3HR-1 line, however, did not vary among the clones tested. No differences in expression of the Epstein-Barr virus-associated nuclear antigen (EBNA) were noted in cells of clones with high or low susceptibility to EA induction. DNA reassociation kinetics demonstrated that Raji cells with high susceptibility to EA induction contained a significantly higher number of Epstein-Barr virus genome equivalents per cell than cells with low susceptibility. Treatment of Raji cells with the tumor promoter TPA did not change the ratio of Epstein-Barr virus-specific DNA to cellular DNA.


No exit: a symbolic interactionist perspective on aging. Most research on socialization for old age, including that of Rosow, Neugarten, and Riley, reflects a normative bias. An alternative perspective on later life is put forth from the symbolic-interactionist tradition in sociology. The concepts of career and status passage reflect an imagery of humans capable of exercising choice and negotiating their way through life. Later life is seen as a unique status passage in that it is inevitable, irreversible, and does not lead to any subsequent status. Controlling the passage therefore assumes great importance.


Tumour antigens and alloantigens. II. Lack of association of rat hepatoma-D23-specific antigen with beta 2 microglobulin. It has previously been shown that the tumour-specific antigen of the chemically-induced rat hepatoma D23 has determinants recognised by alloantisera raised against normal syngeneic liver and spleen. However, no reactions were observed with alloantisera raised against syngeneic erythrocytes suggesting that the alloantigeneic determinant responsible for this cross-reactivity is not a major serologically-defined histocompatibility antigen. This concept has been further examined using a defined turkey anti-rat beta 2 microglobulin antiserum. This antiserum failed to block the binding of syngeneic antihepatoma D23 sera to hepatoma D23 target cells as assessed using membrane immunofluorescence and complement-dependant 51Cr release tests. Furthermore, immune precipitates formed from soluble tumour extracts containing hepatoma D23 specific  antigen with turkey anti-rat beta 2 microglobulin failed to generate a tumour-specific antibody response in syngeneic rats althouth a cross-reactive antiserum was produced following immunization of allogeneic rats with the immune precipitate.


Tumour-related antigen specificities associated with 3-methylcholanthrene-treated rat embryo cells. Rat embryo cells were treated in vitro for 18 h with 10 micrograms/ml 3-methylcholanthrene (MCA). Syngenetic male rats were immunized with several inocula of treated cells to prepare antisera which were screened for membrane immunofluorescence reactivity against panels of established chemically-induced syngeneic rat tumours. Three separate antiserum pools raised against MCA-treated cells reacted with certain chemically-induced tumours, whereas antisera to control (DMSO-treated) cells were completely negative. The reactions observed were reproducible and highly specific for particular target tumours. Absorption studies indicated that each antiserum contained antibodies to several different antigens, present on different tumours. Antiserum prepared against extranuclear membrane from MCA-treated cells, rather than intact MCA-treated cells, was negative. This suggests that the antibody responses were directed against antigens arising subsequently to MCA treatment and injection into syngeneic hosts. It is postulated that carcinomgen treatment results in the acquisition of multiple neoantigens among a treated cell population, which represent an early change in a sequence of events leading to malignant transformation.


The histochemical behaviour, electrophoretic mobility and distribution in cell fractions of acid phosphatase isozymes in prostatic cancer and benign prostate hyperplasia. Acid phosphatase isozymes were investigated in cancerous prostatic tissue (4 cases) and benign prostatic hyperplasia (6 cases). Electron-microscopic histochemical examination of cancer tissue revealed irregular acid beta-glycerophosphatase staining in various cell organelles, including the plasma membrane, which was not seen in non-malignant tissue. Cancerous tissue homogenates also contained isozymic acid phosphatase species with high electrophoretic mobility, which was not detectable in benign tissue unless treated with detergent. Fractionation by differential centrifugation confirmed that much of the acid phosphatase activity in cancer tissue was extra-lysosomal. The detection of these isozyme properties may provide an opportunity, by means of tissue investigations, to define tumour stages earlier than on the basis of increased levels of serum acid phosphatase activity indicative of stage IV (D) prostatic cancer.


Purification of a human prostate specific antigen. Rabbit antiserum raised against the crude extract of normal human prostatic tissue contained antibodies to a prostatic tissue-specific antigen as shown by immunoprecipitation techniques. Using this antiserum a prostate antigen was detected in normal, benign hypertrophic, and malignant prostatic tissues, but not in other human tissues. The prostate antigen was purified to homogeneity from prostatic tissues and showed a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing. This report thus presents the first demonstration of the purification of a prostate-specific antigen that does not represent prostatic acid phosphatase.


[Bone perfusion after plate osteosynthesis. A study of the intact rabbit tibia with disulfin blue vital staining]. After operative fracture treatment an interference with blood supply has to be expected. The fixation of a plate to an intact rabbit tibia lead to extended areas where circulation is disturbed. These disturbances were partially reversible within one week without structural changes visible in the recovering bone. Only the zones remaining without vascular supply after one week started to recover in the 4th week, when intracanalicular remodelling and the formation of new Haversian systems became visible at the borders of the avascular regions.


The effect of clupein on anisotropy and basophilia of polytene chromosomes. When polytene chromosomes are subjected to a clupein treatment, their properties of basophilia and anisotropy are affected. The basophilia is deeply reduced, except in the nucleolar zones, puffs and sites of RNA accumulation. On the other hand, the chromosome birefringence increases. The phenomenon of anomalous dispersion of birefringence usually observed on polytene chromosomes stained with toluidine blue solutions turns into a normal negative dispersion of birefringence, when staining is preceded by clupein treatment. It is concluded that the clupein molecules attach orderly and preferentially to sequential DNA phosphates unbound to chromosome proteins, accentuating DNA anisotropic characteristics. The clupein molecules appear not attaching to RNA phosphates.


Anisotropic staining of apurinic acid with toluidine blue. Using normal rat liver imprints, studies were carried out on the effects of histone extraction and the formation of aldehyde groups from deoxyribose on anisotropic toluidine blue staining of depurinized DNA after sodium bisulfite treatment. The anisotropic effect of bisulfite was found to be determined by binding of bisulfite ions to the aldehyde groups of apurinic acid which, together with free phosphate groups of DNA ensure coparallel attachment of the dye molecules. It was also shown that at pH 5.0 toluidine blue binds with both the phosphate and aldehyde groups of apurinic acid, to give anisotropic staining.


Lung elasticity in regional and diffuse pulmonary fibrosis. Static deflation air and saline pressure-volume (PV) curves were performed on five normal excised baboon lungs, four with radiation-induced upper lobe fibrosis, and two with diffuse bleomycin-induced fibrosis. Curves were compared visually and using half-inflation pressures (h). When plotted as percent of observed vital capacity vs. transpulmonary pressure, the radiation-associated curves were similar in shape to control curves. Also, h values were not significantly different between the two groups. The bleomycin-associated curves, however, were shifted rightward and exhibited increased h values. PV curves were correlated with connective tissue data. Elastin concentration was increased in the irradiated upper lobes through loss of associated constituents. Elastin accumulated in the lower lobes of the irradiated animals through compensatory growth and in the bleomycin-associated lungs through excessive production. Collagen followed a similar but less dramatic trend. We conclude connective tissue accumulation may not lead to rightward shifted PV curves if accumulation is present in relatively noninflatable lung units, or as part of compensatory lung growth.


Descending tracts of the lateral columns of the rat spinal cord: a study using the horseradish peroxidase and silver impregnation techniques. The location of the cells of origin and the projection areas of descending fibre tracts of the spinal cord lateral columns were examined in rats. Unilateral micro-transections of subpopulations of lateral column fibres, at C2 or T10, with subsequent application of horseradish peroxidase to the severed axons, allowed identification, by retrograde labelling, of those cell groups projecting to the spinal cord through the lateral columns. Additionally, the pattern of fibre and preterminal degeneration below the level of transection was examined using the Fink-Heimer silver impregnation technique. The largest number of labelled cells was observed in the ventral portion of nucleus gigantocellularis, projecting ipsilaterally through both the anterolateral (AL) and dorsolateral (DL) columns. Labelled cells were observed in the dorsal portion of the lateral vestibular nucleus (lv) following a T10 transection, and throughout the necleus following a C2 transection. Protein marker was observed in the large Deiters' cells of the lv, ipsilaterally. Also following an AL, but not a DL, column transection, retrograde labelled cells occurred throughout necleus reticularis pontis oralis (rpoo), bilaterally. At the border of rpoo and the lateral lemniscus, a discrete group of labelled cells was observed bilaterally following a DL column transection. This group of reticulospinal cells was located in a position similar to that of the A7 cell group reported in histofluorescence studies. The most extensive group of labelled cells following a DL column transection occurred in the magnocellular portion of the contralateral red nucleus. Although lavelled cells were observed in the red nucleus following either a C2 or T10 DL column transection, labelled cells were more numbeous and extended further rostrally and dorsally, following a high cervical transection. Labelled cells in nucleus raphe magnus were also more numerous following a DL column transection. Additional groups of labelled cells were seen following both an AL or DL column transection. These groups included necleus subcoeruleus ipsilaterally, and nucleus reticularis ventralis and the nucleus of the tract of spinal V, bilaterally. Labelled cells were observed as far forwards as the hypothalamus, occurring predominantly in the paraventricular nucleus, ipsilaterally. A few labelled cells were observed in the lateral hypothalamus. Some cell groups were labelled only after a C2 transection. These included the interstitial nucleus of Cajal, ipsilaterally, the descending vestibular necleus and the deep layers of the superior colliculus, contralaterally, and the central grey matter and nucleus raphe pallidus. Fibre and preterminal degeneration resulting from unilateral AL or DL column transection was examined. Following an AL column transection degeneration was most intense in the ipsilateral laminae V, VI and VII...


Methylmercury hydroxide enhancement of translation and transcription of ovalbumin and conalbumin mRNA's. Translation of total mRNA in heterologous protein-synthesizing systems is often employed as an indirect means of assessing relative mRNA concentrations. However, it is well known that the efficiency of translation of specific mRNAs differs. One such example is the poor translational efficiency of conalbumin mRNA relative to ovalbumin mRNA. In this report we have studied the translation of conalbumin and ovalbumin mRNAs in crude mRNA preparations and with highly purified mRNA preparations. We find that treatment of RNA with methylmercury hydroxide prior to translation improves the translational efficiency of both mRNAs and preferentially improves translational efficiency of conalbumin mRNA to the point where it more correctly reflects the relative concentration of these two mRNAs in crude mRNA preparations. Conalbumin mRNA is also a poor template for the synthesis of full length cDNA synthesis by avian myeloblastosis virus reverse transcriptase, and treatment of this mRNA with methylmercury hydroxide increases the size of DNA sequences synthesized. We conclude that treatment with methylmercury hydroxide produces a partial denaturation of mRNA complexed with either itself or with other RNA molecules and results in more efficient utilization in both translational assays and DNA polymerization reactions.


Heat-induced fragmentation of human alpha 2-macroglobulin. Previous studies have demonstrated that human plasma alpha 2-macroglobulin (alpha 2 M) possesses a single subunit chain (Mr approximately 185,000) when incubated with dodecyl sulfate and dithiothreitol at 37 degrees C and analyzed by dodecyl sulfate-gel electrophoresis. The present study details the observation that heating alpha 2 M to 90 degrees C under identical conditions produces at least two additional polypeptide chains, termed bands II and III, with apparent molecular weights of 125,00 and 62,000. The generation of these fragments is enhanced by increasing the time of incubation. The appearance of band II composition of the buffer, dodecyl sulfate concentrations, or alpha 2 M protein concentration in the incubation mixture. The electrophoretic bands II and III of alpha 2 M have dissimilar 125I-labeled tryptic peptide digests and also differ in their amino acid composition. The heat-induced fragmentation of alpha 2M is not affected by the inclusion of a variety of low molecular weight protease inhibitors, suggesting that the appearance of bands II and III is not due to enzyme-catalyzed hydrolysis. When the subunit chain of alpha 2M is first cleaved by trypsin into the previously described Mr = 85,000 derivative, neither band II nor III material, nor other lower molecular weight products are generated by heat treatment. Furthermore, preincubation of alpha 2M with methylamine prevents fragmentation of the subunit chain. These results indicate that these fragments are neither pre-existing subunits of alpha 2M nor derivatives formed prior to treatment for gel analysis. These data provide evidence that a covalent bond in the alpha 2M molecule is unusually susceptible to heat-induced cleavage.


[Esophago-jejunostomy at the level of the neck for restoration of continuity in carcinoma of the esophagus (after esophageal resection or for palliative reasons) (author's transl)]. A technic of esophago-jejunostomy in the neck is described which uses a Y loop, to which is given a further blood supply through the internal mammary vessels, united to the vessels of the transplanted loop through microsurgical anastomoses. This technic is used either for purely palliative reasons or when the esophageal tumour has become inoperable, or after esophagectomy as a procedure for reconstruction of the digestive tract.


Anaphylatoxin-induced histamine release with human leukocytes: studies of C3a leukocyte binding and histamine release. Purified human C3a and synthetic COOH-terminal peptides of C3a, i.e., a pentapeptide, Leu-Gly-Leu-Ala-Arg (5R), and an octapeptide, Ala-Ala-Ala-Leu-Gly-Leu-Ala-Arg (8R) induced histamine release from human basophil granulocytes. On a molar basis, 5R was one-tenth and 8R was one-fifth as active as C3a in causing histamine release. It was found that 125I-C3a binds to whole leukocytes, interacting with both mononuclear cells and neutrophils and the binding was inhibited by preincubation of cells with unlabeled C3a, but not by C5a. 5R and 8R also inhibited the binding of 125I-C3a to the cells. However, on a molar basis, 2,000 times more 8R or 6,000 times more 5R is required for 50% inhibition of 125I-C3a binding as compared with native C3a. Autoradiography of cells using 125I-C3a and 125I-C5a showed preferential binding of 125I-C3a to eosinophils and basophils, whereas 125I-C5a binds primarily to neutrophils and eosinophils and to a lesser extent to basophils. The preferential binding of C3a and C5a to different cell types may herald significance related to their physiological functions.


Characterization of a distinct nuclear acidic protein antigen (MA) and clinical findings in systemic lupus erythematosus patients with MA antibodies. Circulating antibodies against certain nuclear acidic protein antigens have been shown to have diagnostic and prognostic importance in connective tissue disease. We describe a new precipitin system found in the sera of patients with systemic lupus erythematosus. The antigen, called MA, was prepared from calf thymus nuclei, and was shown to be distinct from other nuclear acidic protein antigens by physicochemical and immunologic techniques. MA antibodies were detected in the serum of 12 of 66 lupus patients and in none of 554 sera from normal controls or patients with other rheumatic diseases. Lupus patients having MA antibodies had more severe disease than did lupus patients with Sm or native DNA antibodies, manifested by recalcitrant skin rashes and a significantly greater incidence of hypocomplementemia, serious renal disease, hypertension, hepatosplenomegaly, lymphadenopathy, and neurological disease (P values range from 0.025 to 0.005). The presence of circulating MA antigen was demonstrated in three lupus patients immediately before a flare of nephritis. These data suggest that MA is a nuclear acidic protein antigen that may identify a subset of lupus patients with very severe disease. The presence of the antigen in the circulation before clinical flares suggests a possible biologic role for the MA system in an immune complex nephritis.


Receptor-mediated binding and internalization of leukocyte elastase by alveolar macrophages in vitro. Radioiodinated leukocyte elastase was found to bind rapidly and specifically to alveolar macrophages in vitro. In contrast to the binding of pancreatic and bacterial proteases, leukocyte elastase binding did not require the presence of alpha 2 macroglobulin. The binding was inhibited by an excess of unlabeled enzyme and was saturable by increasing elastase concentrations. Leukocyte elastase binding thus met criteria for receptor-mediated binding, with and estimated association constant of 4.97 x 10(5) M-1 and an estimated total of 640 x 10(6) binding sites/cell. It differed from the previously described binding of lysosomal glycosidases to macrophages in that it was insensitive to trypsin pretreatment, did not require calcium ions, and was not inhibited by yeast mannan. High-resolution autoradiography indicated that the cell-associated radiolabeled leukocyte elastase was rapidly incorporated into phagolysosomes. Macrophage binding may have a role in clearance of leukocyte elastase from tissue sites where alpha 2 macroglobulin is absent or present in low concentration. Thus, enzyme uptake by alveolar macrophages may be an important factor in the amelioration of lung tissue injury by extracellular leukocyte elastase.


Comparison of spontaneous and experimentally induced canine prostatic hyperplasia. Spontaneous prostatic hyperplasia in the beagle appears to progress with age from a glandular to a cystic histological appearance. Prostatic hyperplasia can be induced in young beagles with intact testes by treatment for 4 mo with either dihydrotestosterone or 5 alpha-androstane-3 alpha, 17 beta-diol, alone, or with either of these steroids in combination with 17 beta-estradiol. In contrast, the induction of prostatic hyperplasia in young castrated beagles, in which the gland had been allowed to involute for 1 mo, requires the administration of both 17 beta-estradiol and either 5 alpha-androstane-3 alpha, 17 beta-diol or dihydrotestosterone. Testosterone and 17 beta-estradiol, either singly or in combination, did not produce the hyperplastic condition in intact or castrated beagles. The experimentally induced prostatic hyperplasia is identical in pathology to the glandular hyperplasia that occurs naturally in the aging dog with intact testes. However, cystic hyperplasia was not produced by any of the treatments tested in young animals.


Acquired lipidosis of marrow macrophages: birefringent blue crystals and Gaucher-like cells, sea-blue histiocytes, and grey-green crystals. Three varieties of compound lipid inclusions occurring as a secondary phenomenon in marrow macrophages are detectable and distinguishable by Romanowsky staining, ultraviolet fluorescence, and polarised light. Birefringent blue crystals and Gaucher-like cells form one variety, sea-blue granules another, and grey-green crystals a third. All occur chiefly in myeloid leukaemias, either acute or chronic.


Alpha-fetoprotein, alpha-1-antitrypsin, and transferrin in gonadal yolk-sac tumours. Since gonadal yolk-sac tumour in pure form or as a component of mixed germ cell tumour is in the majority of patients highly malignant, its histological recognition is of great prognostic importance. Yolk-sac tumour may assume various different histological guises, which have hitherto caused considerable terminological confusion; the present paper is aimed at correlating these morphological diversities with biochemical features which are consistent with yolk-sac differentiation. Using an enzyme-bridge immunoperoxidase technique, a series of 16 gonadal germ cell tumours with a yolk-sac component were screened for the presence of alpha-fetoprotein, alpha-1-antitrypsin, and transferrin. These proteins, normally produced by human yolk sac, were demonstrable in all the morphological patterns of yolk-sac tumour we have previously described. Six malignant non-germ cell tumours were submitted to the same investigations, and no evidence of the three protein markers was found in five; one tumour, however, an oat cell carcinoma of the bronchus, stained positively for transferrin.


A syntax program designed to present base linguistic structures to language-disordered children. An intervention strategy for teaching syntax to language-disordered children is described, and the results of its use in a controlled study with 11 children are reported. The therapy program attempts to move the child from single words through a series of successive approximations to use of the construct noun + "is" + verbing + noun, incorporating auditory directions and visual stimuli in the form of cards and boards upon which the cards are sequentially ordered. The results of the experimental group who participated in this therapeutic sequence were compared to those of 11 control subjects and suggested significant gains as measured by the pre- to postprogram performance (t = 2.31, p 0.05). Advantages and limitations of this type of intervention strategy are also discussed.


The use of visual feedback to control vocal intensity and nasalization. A device which provides visual feedback of vocal intensity, nasalization, or a ratio of nasalization to vocal intensity, is described. The device was used in several experiments in which control of vocal intensity was a necessary component. Normal and hypernasal speakers were able to control their vocal intensities successfully when using the visual display. The display also was used in an experiment designed to determine how well normal speakers can manipulate their nasalization without visual feedback and whether they improve when feedback is provided. Subjects manipulated their nasalization first without feedback, then with feedback, then, again, without feedback. The subjects' performance improved when feedback was provided and determined when feedback was removed. It appears that this device is useful for many applications in a variety of settings.


Piroxicam pharmacokinetics in man: aspirin and antacid interaction studies. Pharmacokinetic studies with piroxicam, a nonsteroidal antiinflammatory agent, have been carried out following the administration of single and multiple oral doses. A plasma half-life of approximately 45 hours is observed, permitting the use of single daily doses in therapy. Enterohepatic recirculation of drug is suggested by the presence of multiple peaks in plasma concentration curves. Piroxicam is highly bound to serum proteins. The absorption and disposition of piroxicam are unaffected by the concomitant administration of aspirin and antiacids. Salicylate plasma levels are similarly unaffected by piroxicam administration.


Transient hypogammaglobulinemia, elevated immunoglobulin E levels, and food allergy. Four infants presented with the combination of food allergy, transient hypogammaglobulinemia (THI), and elevated serum IgE levels. Food allergy was documented by history, positive skin tests for immediate hypersensitivity, radioallergosorbent test, histamine release studies, and lymphocyte transformation in response to food allergens. THI was probably secondary to decreased production since there was no evidence of protein loss from the gastrointestinal tract. Immunologic studies revealed normal B cell number and function in vitro. T cell number and proliferative response to mitogens and antigens were normal but T cells were deficient in their ability to generate helper factors necessary for B cell maturation into immunoglobulin secretory cells. The THI and the deficient production of T cell--helper factor resolved after the age of 20 to 24 mo. A defect in immunoregulation may be responsible for the immunologic abnormalities observed in these patients and their propensity to develop IgE antibodies to food allergens.


Regulation of bowel function by a laxative/stool softener preparation in aged nursing home patients. A 12-week program for 42 chronically constipated patients in a nursing home involved a graduated dosage regimen of standardized senna concentrate with dioctyl sodium sulfosuccinate (Senokot-S Tablets), a high-fiber diet, and an increased fluid intake. The program achieved a satisfactory pattern of bowel evacuation in 36 (86 percent) of these patients. This pattern, as observed during a 4-week follow-up period while therapy was continued, was characterized by absence of fecal impactions, regular comfortable bowel movements of appropriate consistency, minimal or no straining at stool, and minimal or no resort to enemas. In the initial 2-week control period, none of the 42 patients had responded adequately to only dietary modification and increased hydration. Six additional patients whose response to the control regimen was equivocal, also appeared to benefit from the medication program. Two of these were able to discontinue the laxative tablets completely by the end of the 12-week test period. None of the 48 patients who completed the course experienced any adverse effects related to this rehabilitative program.


Occult progressive renal damage in the elderly male due to benign prostatic hypertrophy. A review of the case histories of 345 patients who underwent protatectomy showed that 1.7 percent (6 patients) had "occult and progessive renal damage" secondary to prostatic hypertrophy. All these men were over the age of 60 and the disturbances in micturition were so mild that the patients were unaware of, or chose to ignore them. The presenting symptoms were nonspecific and included generalized weakness, anorexia, nausea, constipation, and weight loss. Investigation revealed impaired renal function of varying degrees. Prostatectomy was associated with a dramatic improvement in all 6 patients. Physicians should be aware of the clinical entity of occult and progressive renal damage secondary to obstruction of the bladder outlet, especially in the elderly male. Uremia can develop with minimal urinary symptoms. Elderly men often suppress or deny their symptoms because of the fear of operation.


Hospitalization of the elderly patient for acute illness. There is a paucity of studies on the characteristics of elderly patients who are admitted for treatment of acute disorders, even though their utilization of hospitals is high. This study involved 828 older persons admitted to a general hospital during a period of six months, for the treatment of acute illness. The typical patient appeared to be a woman in her 70's who entered the medical department because of a cardiovascular or a gastrointestinal disturbance, stayed no longer than 10 days, and returned home to live with her spouse. Though such a person apparently needed minimal assistance in readjusting successfully to the community, there were some exceptions, especially among those who had been living alone or with persons other than a spouse. There was minimal utilization of professional services that might help to assure an optimal post-hospital readjustment.


Neurotoxic effects of leptophos (PhosvelR) in chickens and rats following chronic low-level feeding. Leptophos (O-[4-bromo-2,5 dichlorophenyl] O-methyl phenylphosphonothioate) (PhosvelR) was administered orally to chickens and rats in doses of 0.5 and 5.0 mg/kg/day for 26 weeks. Hens fed 5.0 mg/kg, except one, showed ataxia and became paralysed in the legs at varying times from 8 to 19 weeks. A fifth hen showed ataxia early in the experiment but recovered fully for the remainder of the experiment. Rats fed both doses and chickens fed 0.5 mg/kg showed no signs of delayed neurotoxicity. All hens fed 5.0 mg/kg stopped laying by about the third week. Animals of both species fed 5.0 mg/kg either lost weight (chickens) or gained less weight (rats) than the others. Erythrocyte acetylcholinesterase (AChE) of the chickens given both doses was significantly depressed at first, then increased, and later dropped to control levels. AChE of rats fed 0.5 mg/kg was significantly inhibited but soon recovered to within control levels. On the other hand, the AChE of rats fed 5.0 mg/kg was inhibited throughout the experiment. Plasma cholinesterase (ChE) of both species was first inhibited and then recovered erratically for both insecticide concentrations. Histological alterations in the spinal cord of paralysed hens included axon and myelin degeneration in the ventral, lateral and posterior columns. In the paralysed hens, 79% of the neurotoxic esterase in the brain were inhibited, whereas in the non-paralysed hens (including the one non-paralysed hen receiving 5.0 mg/kg/day) and all rats only about half as much was inhibited.


Immunohistochemical localization of procollagens. I. Light microscopic distribution of procollagen I, III and IV antigenicity in the rat incisor tooth by the indirect peroxidase-anti-peroxidase method. Frozen sections of the growing end of the rat incisor tooth were exposed to antisera or affinity prepared antibodies against partially purified type I, II, or IV procollagen in the hope of detecting the location of the corresponding antigens by the peroxidase-anti-peroxidase technique. The distribution of immunostaining was similar with antisera as with purified antibodies of a given type, but differed for each type; that is, predentin, odontoblasts, pulp and periodontal tissue were the sites of type I; blood vessel walls, pulp and periodontal tissue, of type III; and basement membranes, of type IV antigenicity. It was demonstrated, at least in cases of type I and III, that immunostaining detected the corresponding procollagens and related substances, but not the corresponding collagens. The interpretation of these observations is that: 1) odontoblasts elaborate procollagen I for release to predentin and subsequent transformation to dentinal collagen I; 2) pulp and periodontal cells produce procollagens I and III which presumably become collagens I and III respectively, while the adventitial cells of blood vessels give rise to collagen III; and 3) procollagen IV is associated with basement membranes and, occasionally, adjacent cells.


Immunohistochemical localization of procollagens. II. Electron microscopic distribution of procollagen I antigenicity in the odontoblasts and predentin of rat incisor teeth by a direct method using peroxidase linked antibodies. In an attempt to locate procollagen I in rats odontoblasts, antibodies raised in rabbits were purified by affinity methods and linked to peroxidase. They were then incubated with chopped slices from the growing end of rat incisor teeth. The antibodies binding to the antigens in the slices were visualized by reacting the peroxidase moiety with diaminobenzidine in the presence of hydrogen peroxide. The slices were then embedded in Epon and sectioned for ultrastructural study. Within odontoblasts, the immunostaining indicative of procollagen I antigenicity is moderate in rough endoplasmic reticulum cisternae, strong in spherical and cylindrical Golgi distensions, intense in secretory granules, and variable in lysosomal structures. In predentin, immunostaining is intense close to the odontoblast layer, but decreases gradually in a distal direction. Hence, procollagen I (and/or substances endowed with similar antigenicity such as pro alpha (I) chains and procollagen fragments) is present: 1) along the intracellular pathway of collagen precursors where its concentration gradually increases to reach a maximum in secretory granules; 2) in predentin, into which it is released from the granules for transformation into nonimmunoreactive collagen I; and 3) in lysosomal structures where some of it is hydrolyzed.


A chemical mechanism for tissue staining by osmium tetroxide-ferrocyanide mixtures. The presence of Fe(CN)6(-4) provides sequential, one-electron reduction pathways for OSO4. An equilibrium is established containing OSO4, Fe(CN)6(-4), Fe(CN)6(-3), OSO2(OH)4(-4), and labile cyano-bridged OS-Fe species containing Os in nominal oxidation states of VIII, VII, and VI. These osmium complexes are chelated by appropriately placed donor atoms in the macromolecular tissue matrix, and chelation facilitates the reduction of osmium in situ to lower oxidation states (predominantly IV) that are relatively nonlabile. The greater reactivity and concentration of the Os(VII and VI) intermediates in this system leads to more Os deposition than OsO4 alone; the chelation is responsible for the immobilization of Os and the observed staining pattern in electron micrographs. Chemical data from model systems and electron micrographs of tissue are presented in support of this mechanism.


Nucleolus organizer regions in two species of Bovidae. Silver-staining of nucleolar organizer regions (NORs) in somatic cells of Holstein cattle (Bos taurus) and European buffalo Murrah type (Bubalus bubalus) indicates that the NORs are located in the telomeres of chromosomes 2,3,4,11, and 29 of cattle and chromosomes 3,4,6,23, and 24 of buffalo. Chromosome identification was by Q-banding. Analysis of 326 metaphase cells showed a mean value of 7.7 +/- 1.2 for Ag-NORs with a range between 4 and 10 for cattle. In all of the cells examined, either one or both homologues of chromosome 29 showed telomeric silver staining for cattle. Meiotic preparations showed not more than 5 clusters of silver grains in the pachytene stage of cattle.


Ultrastructural localization of sulfated complex carbohydrates with a modified iron diamine procedure. A thiocarbohydrazide-silver proteinate (TCH-SP) sequence was applied to thin sections of specimens that had been reacted with the high iron diamine (HID) method for ultrastructural localization of sulfated complex carbohydrates. The exposure to TCH-SP enhanced the electron opacity of HID-reactive sites and increased the sensitivity of the procedure. This held true for HID-reacted specimens whether or not they had been post-treated with osmium tetroxide. However in those not postosmicated after HID, the contrast and specificity appeared superior, as sites of osmiophilia were densified equally in specimens exposed to HID, and unexposed controls, by the final osmium tetroxide-TCH-SP sequence. Staining of immature granules of developing polymorphonuclear neutrophils by HID was intensified by the post-treatment with TCH-SP. In addition, granules of blood mononuclear leukocytes and heterophagosomes of peritoneal macrophages revealed HID affinity and hence content of sulfated mucosubstance that was not evident without the TCH-SP steps. Control procedures which entailed initial exposure of the specimen to FeCl3 or MgCl2 solutions and treatment of thin sections with TCH-SP failed to impart density to these sites.


Immunoregulation of localized and disseminated murine myeloma: antigen-specific regulation of MOPC-315 stem cell proliferation and secretory cell differentiation. Tumor development, MOPC-315 stem cells, and M315-secretory cells were quantitated in carrier-primed BALB/c mice that had been challenged subcutaneously or i.v. with mixtures of TNP-carrier and TNP-binding MOPC-315 cells. We observed that tumor incidence, myeloma stem cells, and secretory myeloma cells were: i) suppressed in mice in whom carrier-specific suppressor T cells had previously been induced and ii) initially ehnahced in mice with carrier-specific helper T cells. The early enhancement in mice with carrier-specific helper T cells was followed by progressively declining myeloma stem cell frequencies and regression of established tumors. These studies demonstrate that T cell-derived immunoregulators of host origin can be focused onto localized and disseminated malignant B cells and specifically regulate the expansion and differentiation of the neoplastic clone.


Depletion of NK by cellular immunoadsorption. The binding of human natural killer (NK) cells to their tumor cell targets was investigated by using monolayers of sensitive target cell lines. Monolayers of K562 and HSB, a myeloid and T cell line, respectively, were prepared on poly-L-lysine-coated plastic tissue culture dishes and briefly fixed with 0.2% formaldehyde. Freshly isolated peripheral blood lymphocytes (PBL) were incubated on the monolayers. Nonadherent PBL were then removed, after gentle agitation, by decanting and gently washing the monolayer. They were tested, along with unseparated controls, for NK activity in a short-term 51Cr release assay. PBL that were nonadherent to a tested monolayer had only 20 to 60% of the control cytotoxic activity. Our results suggest that NK recognition sites on the effector lymphocytes were able to interact with reciprocal determinants on the target cell monolayers, resulting in selective loss of NK effector cells from the PBL population. The specificity of the NK effector-target interaction was investigated by testing the ability of each monolayer to remove activity against both targets. These data imply heterogeneity with regard to recognition structure within the NK effector population as well as among the target cells.


Adherent cell function in murine T lymphocyte antigen recognition. III. A macrophage-mediated immune response gene function in the mouse. The I region of the MHC appears to control antigen-specific macrophage-T lymphocyte interaction. The immune response to antigens such as Gl phi 9 are under control of two distinct I subregions, I-A and I-E/I-C. We have asked in a macrophage-dependent, antigen-specific murine T cell proliferation assay whether either or both gene products need be expressed in the antigen-presenting cells. We find that both Ir-Gl phi 9 alpha and beta genes must be expressed and function in the antigen-presenting cell.


Inheritance of tolerance susceptibility to human gamma-globulin in congenic mice. The genetic control of susceptibility to tolerance induction with human gamma-globulin (HGG) was studied by using H-2 congenic mice. Strains tested that were congenic with C57BL/10Sn were completely tolerized by 1.0 mg deaggregated HGG. In contrast A/Sn mice showed full tolerance whereas A.SW mice were only intermediately tolerant. It was further shown that (B10 X SJL)F1 mice could be rendered tolerant but (B10.S X SJL)F1 mice could not. These data indicate a role for H-2 linked genes in control of tolerance susceptibility. Results obtained with the progeny of (B10.S X SJL)F1 backcrossed to B10.S indicate that two non-H-2 linked genes are involved in control of tolerance induction. Preliminary mapping studies show the H-2 gene located to the left of the IC subregion. These results confirm our previous finding that both H-2 and non-H-2 genes control susceptibility of adult mice to tolerance induction with HGG.


Demonstration and partial characterization of 22-nm HBsAg and Dane particles of subtype HBsAg/ady. The present paper describes the demonstration of d, y, w, and r HBsAg determinants in one serum. It was shown that there are two populations of HBsAg particles: HBsAg/ad and HBsAg/ady. All complete Dane particles were of subtype HBsAg/ady. Further characterization of HBsAg/ady particles did not reveal morphologic differences when they were compared with HBsAg/ad and HBsAg/ay particles. An HBsAg/ady phenotype may be the result of a double infection with hepatitis B viruses or exchanges of DNA sequences that determine HBsAg/ay and HBsAg/ad to form a new genotype.


The cytolytic T lymphocyte response to trinitrophenyl-modified syngeneic cells. I. Evidence for antigen-specific helper T cells. Mice were primed subcutaneously with trinitrophenyl (TNP)-modified syngeneic spleen cells. Seven days later, spleen cells from these in vivo primed mice, or spleen cells from naive mice, were co-cultured with TNP-modified syngeneic cells. Spleen cells from the in vivo primed mice demonstrated augmented cytolytic T lymphocyte (CTL) activity. The spleens of these in vivo primed mice contained a population of radioresistant, antigen-specific, helper T cells. Specifically, spleen cells from these mice, after x-irradiation, were able to augment the in vitro CTL response of normal spleen cells to TNP-modified syngeneic cells.


The cytolytic T lymphocyte response to trinitrophenyl-modified syngeneic cells. II. Evidence for antigen-specific suppressor T cells. The subcutaneous administration of trinitrophenyl (TNP)-coupled syngeneic cells 7 days before co-culture with TNP-coupled syngeneic stimulator cells results in increased cytolytic activity. This augmented cytotoxic response has been shown to be dependent, at least partially, on radioresistant "helper" T cells. In this paper we have demonstrated that TNBS-generated suppressor T cells that are capable of suppressing contact sensitivity can specifically suppress the augmented response seen after subcutaneous priming. The i.v. administration of TNP-coupled cells results in priming of the recipient; however, if cells from these animals are transferred to a second recipient, there is evidence of suppressor activity. Thus, the cytotoxic T lymphocyte response is controlled by the same type of complex interactions previously demonstrated for humoral and delayed-type hypersensitivity responses.


Soluble antigen extracts used as blocking agents to obtain specificity in serotyping of Streptococcus mutans. Specific immunofluorescent (IF) conjugates were prepared for Streptococcus mutans serotypes d and g. Serotype specificity was obtained by blocking the cross-reacting antibodies with soluble antigen extracted from cells of the cross-reacting strains. Soluble antigen extracts are particularly useful in obtaining specificity with quantities of conjugates that are too small to be efficiently subjected to adsorption procedures. They also can be used advantageously to block traces of cross-reactivity that frequently remain with conjugates that have been subjected to conventional adsorption procedures. S. mutans isolates from dental plaque samples were identified as belonging to serotypes d or g on the basis of IF staining with conjugates that were made type-specific by blocking the cross-reacting antibodies with appropriate soluble antigen extracts.


A method for specifically detecting internal immunoglobulin by immunofluorescence. The detection of specifically internal immunoglobulin by  staining of fixed preparations of lymphoid cells with appropriate fluorescein-conjugated antisera can be hampered by binding of the conjugates to membrane-bound immunoglobulin. Without preventing staining through membrane-bound immunoglobulin, it is impossible to be sure whether lightly stained cells contain only small amounts of immunoglobulin or wehter the staining is entirely due to the membrane-bound material. Surface immunoglobulin can be stripped from tonsil cells prior to making preparation, and this can be achieved without loss of viable cells. As compared with untreated preparations, pronase-stripped smears contain little non-cellular debris and greatly reduced numbers of small lymphocytes faintly stained by polyvalent anti-human immunoglobulin. The remaining stained cells can be said to specifically contain internal immunoglobulin and are easily scored.


Agarose isoelectric focusing of native human immunoglobulin M and alpha 2-macroglobulin. The resolution of native 19S IgM and alpha 2-macroglobulin by agarose isoelectric focusing is described. The agarose used was practically charge free, thus avoiding disadvantages of electroendoosmosis, and gives a very large network gel, with minimal molecular sieving effects. The resolving power is comparable to that obtained in thin-layer isoelectric focusing in polyacrylamide gel. Clones of human IgM showed a microclonal heterogeneity, similar to IgG antibody heterogeneity, alpha 2-macroglobulin gave a pattern of seven bands in the pI range of 4.1 to 4.9. The flexibility of the agarose isoelectric focusing (IEF) system with regard to immunodetection techniques is illustrated by the use of immunofixation and two-dimensional crossed immunoelectrofocusing. The agarose IEF method has several advantages, viz., non-toxicity, simple handling, uniform and rapid gel formation, and considerably shortened fixing and staining times. The value of the new method is discussed, in particular its usefulness in detecting and isolating IgM antibodies of known specificity produced by cells in culture.


Antigen-specific detection of soluble immune complexes by a solid phase specific antibody system. An antigen-specific immune complex assay based on the following principle has been developed: xenogeneic, antigen-specific antibodies are attached to a solid phase. During first incubation with patient's serum, immune complexes in antigen excess are bound to the xenogeneic antibody by their free antigenic determinants. In a second incubation a labeled antibody, specific for human immunoglobulins, is combined with the antibody part of the immune complex. Quantitation of the label allows the determination of the immune complexes. The principle of the method has been varied using artificial soluble immune complexes of tetanus toxoid and human anti-tetanus toxoid antibody, and immune complexes prepared by mixing patient sera containing either HBsAg or anti-HBsAg antibodies. The reliability of the results is shown by their coefficient of variation (2.5%). With the method described soluble immune complexes predominantly in slight antigen excess, which are thought to be responsible for development of immune complex disease, have been detected.


Cyclic GMP System in epidermis: I. Effect of ischemia. When keratome-sliced pig epidermis was floated on Hank's balanced salt solution, we observed a rapid decrease in the intracellular level of cyclic GMP. A portion of the lost cyclic GMP was detected in the incubation medium. When the epidermis was kept in air at room temperature, the cyclic GMP level also decreased rapidly but to a lesser degree. Incubating the epidermal slice at 37 degrees C in Hank's balanced salt solution with the addition of 3-isobutyl-1-methyl xanthine (IBMX) prevented the decrease. Also, after the cyclic GMP level had fallen, it could be raised to be the in vitro level by the addition of IBMX. Increased amounts of cyclic GMP were detectable in the medium in this case. These data indicate that the decrease in cyclic GMP in ischemic epidermis is due to sudden activation of epidermal cyclic GMP-phosphodiesterase and also in part due to leakage of cyclic GMP extracellularly. In contrast to the rapid decline in the cyclic GMP level, ischemia caused a rapid and transient increase in epidermal cyclic AMP. This confirms previous data by ourselves and by others (Br J Dermatol 92: 249-254, 1975; J Invest Dermatol 68:125-127, 1977). These "ischemic effects" must be avoided in order to measure the "in vivo level" of cyclic nucleotides in epidermis.


Studies on the contact sensitization of man with simple chemicals: V. Clonal priming allows direct in vitro assessment of autologous HLA-associated factors required for immune response to dinitrochlorobenzene. Human lymphocytes from dinitrochlorobenzene (DNCB)-sensitized human subjects primed in first culture with dinitrophenylated-antigens yield a population of cells which respond in an accelerated manner to the same or similar antigen in second culture. Using this "clonal priming" approach, we have demonstrated that such a primed population showed maximal proliferative response to dinitrophenylated autologous cells. These DNP primed clones also showed responses to some dinitrophenylated allogeneic leukocytes. The magnitude of the accelerated blastogenic response with allogeneic leukocytes varied in most instances in relation to the degree of sharing of HLA-A, B, and DRw antigens with the original autologous stimulator. These findings show that the self-specific factors recognized in conjunction with the dinitrophenyl antigen are closely but not invariably associated with established major histocompatibility (MHC)-associated serologic typing results. While DNP primed clones fail to respond to unmodified autologous leukocytes, they often show significant responses to unmodified allogeneic leukocytes. If such accelerated responses to unmodified allogeneic leukocytes are not the result of nonspecific activation of allogeneic responding lymphocyte clones, these findings further indicate that DNP modified self can resemble some alloantigens.


Spatial orientation and distribution of antigens within human glomerular basement membrane. Antibodies to GBM have a linear staining pattern along the GBM by immunofluorescence. In this study, we have examined the distribution of human GBM antigens using Goodpasture antibody and heterologous rabbit anti-human GBM antibody. Goodpasture sera and Goodpasture antibody eluted from diseased kidneys reacted as a single linear component within the GBM. Rabbit anti-human GBM was shown to react along distinctly separate zones of the inner and outer aspects of the GBM; the central portion of the GBM seemed to be nonreactive with this antibody. In comparison, Goodpasture antibodies reacted along the inner zone of the GBM but at a locus external to that reacting with heterologous rabbit anti-GBM antibody. These observations of linear binding of antibody to different sites along the GBM suggest a spatial organization of antigen not previously observed.


XYZ data interpreted by a 12-lead computer program using the derived electrocardiogram. The 12-lead electrocardiogram (ECG) derived from the Frank xyz signals was compared with the conventional 12-lead ECG using the Telemed computer system. In 100 cases studied. Telemed's interpretations were essentially similar in 77, but substantially different in 23. In the 23 cases, interpretations of the derived tracings tended to be more accurate in 14 cases, and less accurate in four cases. In the diagnosis of infarction the probability that the interpretation of the derived tracing will be correct more often was 90%. The better performance may have been related to closer agreement with the vectorcardiogram (VCG). As a substitute for the conventional ECG, the derived ECG offers the prospect of a computerized system that is more practical and more versatile than most currently used systems.


Time course of changes in ventricular excitability and conduction during myocardial ischemia and reperfusion in the dog: effect of lidocaine. Strength-interval curves and conduction times were determined in anesthetized dogs during and following myocardial ischemia using a computerized system capable of determining a 5 point strength-interval curve with conduction times within 20 seconds. At the peak incidence of ligation arrhythmias (5 minutes of ischemia), the falling limb of the strength-interval curve was shifted to the left and conduction time was prolonged, while at 15 minutes of ischemia, the strength-interval was shifted upward and conduction times had returned toward control. Lidocaine enhanced the upward shift of the strength-interval curve, contributing to the electrical stability of the myocardium during this phase of ischemia. During the first minute following abrupt reperfusion of the ischemic zone, there was a slight downward shift of the early part of the strength-interval curve, and conduction times tended to be shorter than control. Lidocaine enhanced the electrophysiological alterations following abrupt reperfusion; that is, it reduced excitation thresholds and increased the tendency to superconductivity. Thus, lidocaine enhanced electrical stability during acute ischemia but tended to exaggerate electrophysiologic defects observed during abrupt reperfusion.


Unusual artifacts in electrocardiographic monitoring. Three unusual artifacts noted during Holter and telemetry monitoring, not previously described, are presented. Recognition of the artifacts prevented misinterpretation and wrong treatment. The clues to the identification of the artifacts and the need for avoiding wrong interpretation and inappropriate treatment are discussed. The cause of the telemetry artifact is discussed.


Atrial repolarization: its role in ST elevation. In four patients, ectopic supraventricular rhythm (or beats) with cephalad anomalous atrial activation and, generally, a short PR, were always associated with an ST elevation in the leads with a negative P wave. An inverted Ta wave appears to be responsible for the ST elevation.


Acid fast staining of oxidized neuromelanin and lipofuscin in the human brain. Acid fast staining of the bleached residuum of substantia nigra neuromelanin and of oxidized inferior olive lipofuscin was demonstrated in paraffin and frozen sections stained with the acetic acid, carbol fuchsin method of Barbeito-Lopez and the aldehyde fuchsin method of Gomori. Acid fast staining occurred when sections were exposed to a prestain oxidation with potassium permanganate in conjunction with a poststain differentiation with dilute hydrochloric acid. The acid fast staining with acetic acid, carbol fuchsin was differentiable and in contrast to the acid fast staining with aldehyde fuchsin which was nondifferentiable. A possible histochemical basis for differentiable and nondifferentiable acid fast staining was discussed. The identify of the bleached residuum of neuromelanin as lipofuscin was also discussed.


Definition of cellular immune responses to brain antigens in human head trauma. Cellular immune responses to brain antigens in patients with head injury were studied by applying the leukocyte adherence inhibition (LAI) assay. The investigation was conducted in three phases. 1) In the initial phase, evaluation of a series of 22 test and 25 control cases obtained at random during a 2- to 6-week time frame following a traumatic event indicated significant non-adherence of leukocytes (NAL) in 77% of the test group and 20% of the control group in the presence of brain antigen. 2) In a second phase, larger test population was divided into four groups of different posttraumatic intervals. This study measured NAL in the presence of normal heart of normal brain antigen. Assays revealed an initial significant NAL in the presence of both antigens; however, after the first week following injury the majority of cases manifested significant NAL only with brain antigen. These values of NAL persisted over a 6- to 8-week period. 3) As a final phase of investigation, analysis of a sequential series of assays in 12 patients over a 90-day period indicated significant NAL in the presence of brain antigen within the first week of injury, this was followed by a drop in NAL in most of the cases. Studies at 7 to 60 days posttrauma demonstrated significant NAL with brain antigen alone, with a subsequent drop by 90 days. These observations are interpreted to represent sensitization of leukocyte subgroups to brain proteins that are immunologically recognized following the traumatic event.


Alphafetoprotein and human chorionic gonadotropin determination in cerebrospinal fluid. An aid to the diagnosis and management of intracranial germ-cell tumors. The cerebrospinal fluid (CSF) and serum of six patients with histologically verified intracranial germ-cell tumors were assayed serially for the presence of alphafetoprotein (AFP) and the beta subunit of human chorionic gonadotropin (HCG). Two patients had embryonal carcinomas, two had choriocarcinomas, and two had dysgerminomas. The marker profile for a given tumor in either CSF or serum correlated with the histological diagnosis; that is, embryonal carcinoma produced AFP and HCG, choriocarcinoma produced HCG, and dysgerminoma produced no markers. The marker levels in serum and CSF declined with therapy and rose usually prior to the development of overt clinical symptoms if the patient's tumor recurred. A CSF-to-serum gradient of the marker levels was present in three of four patients, and the serum levels were often normal when the CSF values were elevated. Ventricular marker levels were lower than the lumbar levels in two of two patients. The assay of these biological markers is a sensitive indicator of the success of therapy, and the presence of a CSF-to-serum gradient suggests that the major portion of the neoplasm rests within the central nervous system. A histological diagnosis can be inferred without the necessity of surgery in appropriate clinical contexts.


Antigenic subtypes of HBsAg: their distribution and pattern of occurence among blood donors and patients with liver diseases and leprosy in Tamilnadu, India. One hundred and seventy hepatitis B Surface antigen positive sera derived from blood donors, patients with liver diseases and leprosy were antigenically subtyped by Rheophoresis and 107 of them by agar-gel diffusion. For the first time in India HBsAg/adr as a predominant subtype (64.0%) is documented. Of the two methods adopted, Rheophoresis showed a greater sensitivity of typing, namely 82.3% in contrast to 39.2% only by agar-gel diffusion (p less than 0.001). Analysis of the Hepatitis Be antigen and antibody (anti HBe) positive sera for subtype predeliction revealed the same pattern as in HBe system negative sera.


Etiology of human liver cancer: controlled prospective study in liver cirrhosis. The incidence of primary liver cell carcinoma was investigated in a prospective study over 6 yr and 5 mo in 403 clinically unselected patients derived from a homogeneous population by means of serial determination of alpha 1-fetoprotein (AFP) by radioimmunoassay. The diagnosis of liver cirrhosis was proved in 90% by laparoscopy and/or histology and/or autopsy. The incidence of primary liver cell carcinoma in liver cirrhosis in the clinically studied patients was 4.47%, significantly lower than in the autopsy material (11.03%; p less than or equal to 0.025). In the follow-up study, all patients with increasing AFP concentrations exhibited a primary liver cell carcinoma. A transitory rise of AFP (higher than 50 ng/ml) was observed in 15.1% of patients with liver cirrhosis without primary liver cell cancer. In contrast to the results of animal experiments, this transitory rise of AFP was not followed by malignant transformation of the cirrhotic tissue. Posthepatitic liver cirrhosis was observed in 21.57%, postalcoholic liver cirrhosis in 42.93%, and cryptogenic liver cirrhosis in 27.30%. Liver cirrhosis of other etiology occurred in 8.19%. The incidences of primary liver cell cancer in these 4 groups were 4.94, 4.62, 5.45, and 0%, respectively. These differences are not statistically significant, although in absolute figures postalcoholic liver cirrhosis is the main cause of primary liver cell carcinoma in this sample from West Germany. HBs antigen-positive liver cirrhosis was more often associated with primary liver cell cancer than HBs antigen-negative liver cirrhosis (6.58 versus 3.96%); this difference also is not statistically significant. Observations of larger groups of patients may show a higher risk of developing primary liver cell carcinoma in those with a combination of alcohol abuse and HBs antigenemia and/or acute hepatitis in the history. Patients without these 2 risk factors had an incidence of primary liver cell carcinoma of 2.61%; those with 1 risk factor, 5.77%; and those with both risk factors, 10.71%.


Clinical aspects of alpha 1-fetoprotein determination in human liver cancer and in humans and experimental animals at carcinogenic risk. Recent findings concerning the significance of alpha 1-fetoprotein (AFP) as a tool for clinical diagnosis and monitoring of tumor diseases are reviewed briefly. The applicability of this protein marker to the early diagnosis of patients at carcinogenic risk is discussed. In addition, experimental data obtained with a model of chemical hepatocarcinogenesis are reported. The increase of proliferative activity in precancerous liver tissue preceded AFP production under experimental conditions with azodyes and aflatoxin B1 as carcinogens. Immunohistochemical analysis of the relation of AFP to changes of cell populations and to liver tissue rearrangement led to the conclusion that AFP-producing cells cannot be precursors of malignant hepatocytes; however, AFP appeared to be linked to dividing hepatocytes at a certain step of cell differentiation regardless of the stages of precancerous development. A decrease in the rate of nuclear RNA synthesis was observed in both precancerous and tumor tissues. A possible analogy between the early phase of AFP production in animal carcinogenesis and that in human carcinogenesis is considered.